Anti-il36r antibodies

ABSTRACT

The present invention provides antibodies and antigen-binding fragments (e.g., human E antibodies) that bind specifically to human IL-36 receptor. Methods for treating or preventing diseases mediated by IL36R (e.g., skin or colon inflammatory conditions such as palmo-plantar pustular psoriasis, palmoplantar pustulosis, generalized pustular psoriasis, ulcerative colitis or IBD) using the antibodies and fragments are also provided along with methods of making the antibodies and fragments.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 62/698,482, filed Jul. 16, 2018; U.S. Provisional PatentApplication No. 62/846,989, filed May 13, 2019; and U.S. ProvisionalPatent Application No. 62/866,028, filed Jun. 25, 2019; each of which isherein incorporated by reference in its entirety.

FIELD OF THE INVENTION

The field of the invention relates, in part, to antibodies that bind toIL-36 receptor and the use of such antibodies to treat inflammatorydisorders including psoriasis or inflammatory bowel disease.

INCORPORATION BY REFERENCE OF SEQUENCE LISTING

The Sequence Listing in the ASCII text file, named as36432_10484US01_SequenceListing of 176 KB, created on Jul. 15, 2019 andsubmitted to the United States Patent and Trademark Office via EFS-Web,is incorporated herein by reference.

BACKGROUND OF THE INVENTION

The interleukin (IL)-36 cytokines include 3 agonists, IL-36α, IL-36β,and IL-36γ that bind to a common receptor composed of IL-36R andIL-1RAcP to stimulate inflammatory responses. IL-36 receptor (IL-36R) isa single-pass membrane receptor for a subset of the IL-1 family ofcytokines, IL-36α, IL-36β, and IL-36γ, and upon binding to any of theseligands, there is recruitment of its co-receptor, the IL-1R accessoryprotein (IL-1RAcP), which induces a signaling cascade that involves NFκBand mitogen-activated kinase pathways (Sims et al, 2010.

A mediator of some inflammatory skin conditions, such as psoriasis, isIL-36. Psoriasis is a common, immune-mediated, inflammatory skin diseasewhich includes the variants plaque psoriasis and generalized pustularpsoriasis. Standard therapeutic guidelines include the use of topicalsteroids, topical vitamin D, systemic immunosuppressants and variousbiologics, such as anti-tumor necrosis factor (TNF) α, anti-interleukin(IL)-23 and anti-IL-17 antibodies. IL-36 members are overexpressed inthe lesional skin of plaque psoriasis and activation of IL-36R mightcontribute to the persistence and perpetuation of psoriatic inflammationtogether with the TNF-α/IL-23/IL-17/IL-22 axis. (Di Cesare et al., TheIL-23/Th17 axis in the immunopathogenesis of psoriasis, Journal ofInvestigative Dermatology 129: 1339-1350 (2009) and Blumberg et al.,IL-1RL2 and its ligands contribute to the cytokine network in psoriasis.J Immunol 185: 4354-4362 (2010)).

Currently available treatments for palmoplantar pustulosis (PPP) andpalmoplantar pustular psoriasis (PPPP), however, are limited. Spesolimaband ANB019 are anti-IL36R antibodies in clinical development whichsuffer from drawbacks related to immunogenicity and potency.

SUMMARY OF THE INVENTION

The present invention provides anti-IL36R antibodies and antigen-bindingfragments thereof that exhibit superior properties. For example, weobserved in pharmacokinetic studies in three cynomolgous monkeys pergroup (0.5 and 5 mg/kg subcutaneous dose groups; n=3/group), that theanti-IL36R antibodies set forth herein (e.g., H4H14708P2) exhibitedabout 1.2-fold greater exposure than anti-IL36R antibody, APE6155.Moreover, we also observed that APE6155 exhibited less potency thananti-IL36R antibodies set forth herein, e.g., in reducing skin thicknessand pathology scores in IMQ-induced skin inflammation and in reducingpro-inflammatory cytokines in skin. Spesolimab, a humanized anti-IL36Rantibody, exhibited high levels of anti-drug antibody in human subjectswith GPP. In a Phase 1 clinical trial, 3 of 7 patients had anti-drugantibodies at week 2 and these sustained to week 20 after a single dose.This property of spesolimab would not be ideal for chronic long-termtreatment. Amin, First Data in GPP from Competitor Anti-IL36 ProvidesProof of Concept of ANB019, Flash Note, Company Update, AnaptysBio,Jefferies (Sep. 16, 2018). The human anti-IL36R antibodies of thepresent invention are not expected to be highly immunogenic in humans.

The present invention provides an antigen-binding protein (e.g., anantibody or antigen-binding fragment thereof, e.g., a human antibody orantigen-binding fragment thereof or a multispecific antibody) that (i)specifically binds to the same epitope on IL36R as a referenceantigen-binding protein; or (ii) competes for binding to IL36Rpolypeptide with a reference antigen-binding protein, wherein thereference antigen-binding protein comprises: (a) a heavy chainimmunoglobulin that comprises CDR-H1, CDR-H2 and CDR-H3 of a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184,188, 192, 196, 200, 204, 208, 212, 216, 220 or 224; and/or (b) a lightchain immunoglobulin that comprises CDR-L1, CDR-L2 and CDR-L3 of a lightchain immunoglobulin that comprises the amino acid sequence set forth inSEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190,194, 198, 202, 206, 210, 214, 218, 222 or 226. For example, in anembodiment of the invention, the antigen-binding protein comprises: (i)a heavy chain immunoglobulin that comprises CDR-H1, CDR-H2 and CDR-H3 ofa heavy chain immunoglobulin that comprises the amino acid sequence setforth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170,180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or 224; and/or(ii) a light chain immunoglobulin that comprises CDR-L1, CDR-L2 andCDR-L3 of a light chain immunoglobulin that comprises the amino acidsequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146,162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or 226. In anembodiment of the invention, the antigen-binding protein comprises (a) aheavy chain immunoglobulin variable region comprising an amino acidsequence having at least 90% amino acid sequence identity to the aminoacid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114,130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216,220 or 224; and/or (b) a light chain immunoglobulin variable regioncomprising an amino acid sequence having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 10,26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202,206, 210, 214, 218, 222 or 226. For example, in an embodiment of theinvention, the antigen-binding protein comprises: (a) a heavy chainimmunoglobulin comprising the CDR-H1, CDR-H2 and CDR-H3 of a heavy chainimmunoglobulin comprising an amino acid sequence set forth in SEQ ID NO:2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184, 188, 192,196, 200, 204, 208, 212, 216, 220 or 224 and at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 2,18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184, 188, 192,196, 200, 204, 208, 212, 216, 220 or 224; and/or (b) an light chainimmunoglobulin comprising the CDR-L1, CDR-L2 and CDR-L3 of a light chainimmunoglobulin comprising an amino acid sequence set forth in SEQ ID NO:10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198,202, 206, 210, 214, 218, 222 or 226 and at least 90% amino acid sequenceidentity to the amino acid sequence set forth in SEQ ID NO: 10, 26, 42,58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210,214, 218, 222 or 226. In an embodiment of the invention, theantigen-binding protein comprises: a heavy chain immunoglobulin thatcomprises: CDR-H1 comprising the amino acid sequence set forth in SEQ IDNO: 4; CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:6; and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:8 and/or CDR-H1 comprising the amino acid sequence set forth in SEQ IDNO: 20; CDR-H2 comprising the amino acid sequence set forth in SEQ IDNO: 22; and CDR-H3 comprising the amino acid sequence set forth in SEQID NO: 24 and/or CDR-H1 comprising the amino acid sequence set forth inSEQ ID NO: 36; CDR-H2 comprising the amino acid sequence set forth inSEQ ID NO: 38; and CDR-H3 comprising the amino acid sequence set forthin SEQ ID NO: 40 and/or CDR-H1 comprising the amino acid sequence setforth in SEQ ID NO: 52; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 54; and CDR-H3 comprising the amino acid sequenceset forth in SEQ ID NO: 56 and/or CDR-H1 comprising the amino acidsequence set forth in SEQ ID NO: 68; CDR-H2 comprising the amino acidsequence set forth in SEQ ID NO: 70; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 72 and/or CDR-H1 comprising theamino acid sequence set forth in SEQ ID NO: 84; CDR-H2 comprising theamino acid sequence set forth in SEQ ID NO: 86; and CDR-H3 comprisingthe amino acid sequence set forth in SEQ ID NO: 88 and/or CDR-H1comprising the amino acid sequence set forth in SEQ ID NO: 100; CDR-H2comprising the amino acid sequence set forth in SEQ ID NO: 102; andCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 104and/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:116; CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:118; and CDR-H3 comprising the amino acid sequence set forth in SEQ IDNO: 120 and/or CDR-H1 comprising the amino acid sequence set forth inSEQ ID NO: 132; CDR-H2 comprising the amino acid sequence set forth inSEQ ID NO: 134; and CDR-H3 comprising the amino acid sequence set forthin SEQ ID NO: 136 and/or CDR-H1 comprising the amino acid sequence setforth in SEQ ID NO: 140; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 142; and CDR-H3 comprising the amino acid sequenceset forth in SEQ ID NO: 144 and/or CDR-H1 comprising the amino acidsequence set forth in SEQ ID NO: 156; CDR-H2 comprising the amino acidsequence set forth in SEQ ID NO: 158; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 160 and/or CDR-H1 comprising theamino acid sequence set forth in SEQ ID NO: 172; CDR-H2 comprising theamino acid sequence set forth in SEQ ID NO: 174; and CDR-H3 comprisingthe amino acid sequence set forth in SEQ ID NO: 176 and/or CDR-L1comprising the amino acid sequence set forth in SEQ ID NO: 12; CDR-L2comprising the amino acid sequence set forth in SEQ ID NO: 14; andCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 16and/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:28; CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:30; and CDR-L3 comprising the amino acid sequence set forth in SEQ IDNO: 32 and/or CDR-L1 comprising the amino acid sequence set forth in SEQID NO: 44; CDR-L2 comprising the amino acid sequence set forth in SEQ IDNO: 46; and CDR-L3 comprising the amino acid sequence set forth in SEQID NO: 48 and/or CDR-L1 comprising the amino acid sequence set forth inSEQ ID NO: 60; CDR-L2 comprising the amino acid sequence set forth inSEQ ID NO: 62; and CDR-L3 comprising the amino acid sequence set forthin SEQ ID NO: 64 and/or CDR-L1 comprising the amino acid sequence setforth in SEQ ID NO: 76; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 78; and CDR-L3 comprising the amino acid sequenceset forth in SEQ ID NO: 80 and/or CDR-L1 comprising the amino acidsequence set forth in SEQ ID NO: 92; CDR-L2 comprising the amino acidsequence set forth in SEQ ID NO: 94; and CDR-L3 comprising the aminoacid sequence set forth in SEQ ID NO: 96 and/or CDR-L1 comprising theamino acid sequence set forth in SEQ ID NO: 108; CDR-L2 comprising theamino acid sequence set forth in SEQ ID NO: 110; and CDR-L3 comprisingthe amino acid sequence set forth in SEQ ID NO: 112 and/or CDR-L1comprising the amino acid sequence set forth in SEQ ID NO: 124; CDR-L2comprising the amino acid sequence set forth in SEQ ID NO: 126; andCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 128and/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:124; CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO:126; and CDR-L3 comprising the amino acid sequence set forth in SEQ IDNO: 128 and/or CDR-L1 comprising the amino acid sequence set forth inSEQ ID NO: 148; CDR-L2 comprising the amino acid sequence set forth inSEQ ID NO: 150; and CDR-L3 comprising the amino acid sequence set forthin SEQ ID NO: 152 and/or CDR-L1 comprising the amino acid sequence setforth in SEQ ID NO: 164; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 166; and CDR-L3 comprising the amino acid sequenceset forth in SEQ ID NO: 168 and/or CDR-L1 comprising the amino acidsequence set forth in SEQ ID NO: 124; CDR-L2 comprising the amino acidsequence set forth in SEQ ID NO: 126; and CDR-L3 comprising the aminoacid sequence set forth in SEQ ID NO: 128. In an embodiment of theinvention, the antigen-binding protein comprises: (1) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 4; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 6; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 8; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 12; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 14; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 16; (2) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 20: a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 22; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 24; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 28; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 30; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 32; (3) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 36; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 38; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 40; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 44; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 46; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 48; (4) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 52; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 54; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 56; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 60; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 62; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 64; (5) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 68; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 70; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 72; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 76; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 78; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 80; (6) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 84; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 86; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 88; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 92; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 94; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 96; (7) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 100; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 102; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 104; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 108; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 110; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 112; (8) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 116; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 118; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 120; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 124; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 126; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 128; (9) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 132; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 134; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 136; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 124; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 126; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 128; (10) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 140; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 142; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 144; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 148; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 150; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 152; (11) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 156; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 158; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 160; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 164; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 166; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 168; or (12) a heavy chainimmunoglobulin variable region comprising a CDR-H1 comprising the aminoacid sequence set forth in SEQ ID NO: 172; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 174; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 176; and a light chainimmunoglobulin variable region comprising a CDR-L1 comprising the aminoacid sequence set forth in SEQ ID NO: 124; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 126; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 128. In an embodiment of theinvention, the antigen-binding protein comprises (a) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184,188, 192, 196, 200, 204, 208, 212, 216, 220 or 224; and/or (b) a lightchain immunoglobulin that comprises the amino acid sequence set forth inSEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190,194, 198, 202, 206, 210, 214, 218, 222 or 226. The present inventionincludes an antigen-binding protein (e.g., antibody or antigen-bindingfragment thereof) comprising: (a) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO: 2,and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 10; (b) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 18, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:26; (c) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 34, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 42; (d) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:50, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 58; (e) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 66, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:74; (f) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 82, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 90; (g) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:98, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 106; (h) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 114, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:122; (i) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 130, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 122; (j) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:138, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 146; (k) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 154, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:162; and/or (l) a heavy chain immunoglobulin variable region thatcomprises the amino acid sequence set forth in SEQ ID NO: 170, and alight chain immunoglobulin variable region that comprises the amino acidsequence set forth in SEQ ID NO: 122—for example wherein the heavy chainimmunoglobulin variable region is linked to an heavy chain constantregion (e.g., IgG (e.g., IgG1 or IgG4)) and the light chainimmunoglobulin variable region is linked to a light chain constantregion (e.g., lambda or kappa). For example, the light and heavy chainconstant regions are human constant regions. In an embodiment of theinvention, the antigen-binding protein (e.g., antibody orantigen-binding fragment thereof) of the present invention comprises:(a) a heavy chain immunoglobulin that comprises the amino acid sequenceset forth in SEQ ID NO: 180, and a light chain immunoglobulin thatcomprises the amino acid sequence set forth in SEQ ID NO: 182; (b) aheavy chain immunoglobulin that comprises the amino acid sequence setforth in SEQ ID NO: 184, and a light chain immunoglobulin that comprisesthe amino acid sequence set forth in SEQ ID NO: 186; (c) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 188, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 190; (d) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 192, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 194; (e) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 196, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 198; (f) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 200, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 202; (g) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 204, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 206; (h) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 208, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 210; (i) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 212, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 214; (j) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 216, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 218; (k) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 220, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 222; and/or (l) a heavy chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 224, and a light chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 226.

Antigen-binding proteins of the present invention, may, in an embodimentof the invention, be characterized by one or more of the followingproperties:

-   -   Binds to human IL36R with a K_(D) of about 2.18 nM to about 13.9        nM at 25° C. or with a K_(D) of about 4.25 nM to about 29.5 nM        at 37° C.;    -   Binds to Macaca fascicularis IL36R with a K_(D) of about 7.87 nM        to about 34.4 nM at 25° C. or with a K_(D) of about 14.4 nM to        about 58.2 nM at 37° C.;    -   Binds to human IL36R fused to a mouse IgG2a with a K_(D) of        about 173 pM to about 5.79 nM at 25° C. or with a K_(D) of about        205 pM to about 28.7 nM at 37° C.;    -   Binds to human IL36R fused to IL1RAcP extracellular domain        expressed with mouse IgG2a Fc tag with a K_(D) of about 212 pM        to about 14 nM at 25° C. or with a K_(D) of about 264 pM to        about 40.9 nM at 37° C.;    -   Competes with H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;        H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2;        H4H14757P; H4H14758P or H4H14760P2 for binding to IL36R;    -   Blocks activation of one or more NFκB elements, which is/are        fused to a reporter gene, in a host cell, by IL-36R (e.g., human        or Macaca fascicularis) in the presence of IL-1RAcP and IL36R        ligand;    -   Prevents or ameliorates skin inflammation or reduces skin        thickness or total pathology score or reduces pro-inflammatory        cytokine levels in a subject suffering from skin inflammation;    -   Prevents or ameliorates colitis or colon inflammation or reduces        fecal levels of LCN2 polypeptide in a subject with such colitis        or inflammation;    -   Protects residues (a) 113-119, 113-122, 116-119 and/or 116-122;        and/or (b) 264-271, 267-271, 268-271, 268-276, 268-277 and/or        271-276, of human IL36R (IL-1 RL2) set forth herein in SEQ ID        NO: 227 (or the corresponding residues in wild-type IL-1RL2),        when bound, from digestion with pepsin and/or Protease XIII        and/or deuteration in the presence of deuterium;    -   Binds to IL36R (IL-1RL2) (e.g., human IL36R) at residues        113-119, 113-122, 116-119, 116-122, 264-271, 267-271, 268-271,        268-276, 268-277 and/or 271-276 of human IL36R comprising the        amino acid sequence set forth herein in SEQ ID NO: 227 (or the        corresponding residues in wild-type IL-1RL2);    -   Binds Domain II of IL36R (IL-1RL2) (e.g., human IL36R), e.g.,        with a coverage of about 80.0, 80.1, 81.0 or 81.5% or about        80-81 or 80-82% coverage; and/or    -   Binds a polypeptide comprising the amino acid sequence        YKQILHLGKD (SEQ ID: 229) (amino acids 113-122 of SEQ ID NO:        227):    -   Inhibits IL36a, IL36 and/or IL36γ (e.g., at a concentration of        about 10 nM), e.g., in in vitro epidermal keratinocytes,        intestinal myofibroblasts and/or CD14+ monocytes, with an IC₅₀        of about 1, 2, 3, 4, 5 or 6 nM or 1-6 nM; and/or    -   Competitively inhibits IL36a, IL36 and/or IL36γ-mediated        activation of NFκB (e.g., an NFκB response element        (5×)-luciferase-IRES-GFP reporter in a cell such as HEK293) by        IL36R; for example, as measured in a Schild Assay format.

Complexes comprising an IL36R polypeptide or antigenic fragment thereofcomplexed with an antigen-binding protein of the present invention(e.g., an antibody or antigen-binding fragment thereof, e.g., a humanantibody or antigen-binding fragment thereof or a multispecificantibody) are also within the scope of the present invention.

Also provided by the present invention are methods for making anantigen-binding protein of the present invention (e.g., an antibody orantigen-binding fragment thereof, e.g., a human antibody orantigen-binding fragment thereof or a multispecific antibody) or animmunoglobulin chain thereof comprising: (a) introducing one or morepolynucleotides encoding an immunoglobulin chain of said antigen-bindingprotein into a host cell (e.g., a Chinese hamster ovary (CHO) cell); (b)culturing the host cell under conditions favorable to expression of thepolynucleotide; and (c) optionally, isolating the antigen-bindingprotein or immunoglobulin chain from the host cell and/or medium inwhich the host cell is grown. Antigen-binding proteins andimmunoglobulin chains which are products of such a method are also partof the present invention.

The present invention also provides a polypeptide comprising: (a)CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain variableregion of an immunoglobulin chain that comprises the amino acid sequenceset forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154,170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or 224;and/or (b) CDR-L1, CDR-L2, and CDR-L3 of immunoglobulin light chainvariable region of an immunoglobulin chain that comprises the amino acidsequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146,162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or 226; or,any amino acid sequence set forth herein, e.g., (c) the amino acidsequence set forth in a member selected from the group consisting of SEQID NO: 1-226. A polynucleotide encoding one or more (e.g., 2, e.g., aheavy and a light chain immunoglobulin set forth herein) of suchpolypeptides are also part of the present invention. Vectors, e.g.,plasmids, comprising such a polynucleotide are also part of presentinvention. A host cell (e.g., a CHO cell) comprising any antigen-bindingprotein or immunoglobulin chain or polypeptide or polynucleotide orvector set forth herein is part of the present invention, e.g., whereinthe polynucleotide and/or vector is integrated into a chromosome of thehost cell or is ectopic.

A composition or kit comprising one or more of the antigen-bindingproteins set forth herein (e.g., an antibody or antigen-binding fragmentthereof, e.g., a human antibody or antigen-binding fragment thereof or amultispecific antibody), optionally in association with a furthertherapeutic agent (e.g., an anti-inflammatory agent, an anti-TNFalphaantibody or antigen-binding fragment thereof, an IL17 inhibitor, anIL23p19 inhibitor, an IL12p40 inhibitor, guselkumab, ustekinumab,brodalumab, ixekizumab, secukinumab, one or more human TNF receptors orfragments thereof linked to an immunoglobulin, infliximab, adalimumab,etanercept, dupilumab, sarilumab, tocilizumab, golimumab, abatacept,tofacitinib, abatacept, a non-steroidal anti-inflammatory drug (NSAID),ibuprofen, naproxen, acetaminophen, aspirin, celecoxib,cyclophosphamide, methotrexate, a corticosteroid, cortisone andprednisone, form part of the present invention.

Pharmaceutical compositions comprising an antigen-binding protein setforth herein (e.g., an antibody or antigen-binding fragment thereof,e.g., a human antibody or antigen-binding fragment thereof or amultispecific antibody) and a pharmaceutically acceptable carrier and,optionally, a further therapeutic agent, are also part of the presentinvention.

The present invention also provides a vessel or injection device (e.g.,a pre-filled syringe) comprising an antigen-binding protein (e.g., anantibody or antigen-binding fragment thereof, e.g., a human antibody orantigen-binding fragment thereof or a multispecific antibody) orcomposition set forth herein.

The present invention further provides a method for treating orpreventing an IL36R mediated disorder (e.g., an inflammatory orautoimmune disease or inflammatory bowel disease) in a subject in needthereof (e.g., a human), comprising administering (e.g., parenterally),to the subject, a therapeutically effective amount of antigen-bindingprotein as set forth herein (e.g., an antibody or antigen-bindingfragment thereof, e.g., a human antibody or antigen-binding fragmentthereof or a multispecific antibody), optionally in association with afurther therapeutic agent (e.g., an anti-inflammatory agent).

The present invention also provides a method for administering anantigen-binding protein as set forth herein (e.g., an antibody orantigen-binding fragment thereof, e.g., a human antibody orantigen-binding fragment thereof or a multispecific antibody) into thebody of a subject (e.g., a human) comprising injecting (e.g.,subcutaneously, intravenously or intramuscularly) the antigen-bindingprotein into the body of the subject, optionally in association with afurther therapeutic agent (e.g., an anti-inflammatory agent).

The present invention encompasses any polypeptide comprising an aminoacid sequence which is set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14,16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50,52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86,88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116,118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144,146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172,174, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220, 224,182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 and/or 226 or avariant thereof.

The present invention includes any polynucleotide comprising anucleotide sequence which is set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11,13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47,49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83,85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115,117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143,145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171,173, 175, 179, 183, 187, 191, 195, 199, 203, 207, 211, 215, 219, 223,181, 185, 189, 193, 197, 201, 205, 209, 213, 217, 221 and/or 225 or avariant thereof.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Sequence comparison between germline and V_(H) and V_(L) OfH4H14706P2.

FIG. 2. Sequence comparison between germline and V_(H) and V_(L) OfH4H14708P2.

FIGS. 3A-3F. Increasing concentration of H4H14706P2 and H4H14708P2generated rightward shift of IL-36α (3A and 3D), IL-36β (3B and 3E), orIL-36γ (3C and 3F) dose response curves revealing competitive nature ofinhibition by H4H14706P2 and H4H14708P2 (RLU, relative light units)

FIG. 4. H4H141708P2 and APE6155 pharmacokinetic analysis (concentrationof antibody in serum over time) in cynomolgus monkeys dosedsubcutaneously with 0.5 mg/kg or 5.0 mg/kg of antibody.

DETAILED DESCRIPTION OF THE INVENTION

In accordance with the present invention there may be employedconventional molecular biology, microbiology, and recombinant DNAtechniques within the skill of the art. Such techniques are explainedfully in the literature. See, e.g., Sambrook, Fritsch & Maniatis,Molecular Cloning: A Laboratory Manual, Second Edition (1989) ColdSpring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (herein“Sambrook, et al., 1989”); DNA Cloning: A Practical Approach, Volumes Iand II (D. N. Glover ed. 1985); Oligonucleotide Synthesis (M. J. Gaited. 1984); Nucleic Acid Hybridization (B. D. Hames & S. J. Higgins eds.(1985)); Transcription And Translation (B. D. Hames & S. J. Higgins,eds. (1984)); Animal Cell Culture (R. I. Freshney, ed. (1986));Immobilized Cells And Enzymes (IRL Press, (1986)); B. Perbal, APractical Guide To Molecular Cloning (1984); F. M. Ausubel, et al.(eds.), Current Protocols in Molecular Biology, John Wiley & Sons, Inc.(1994).

An anti-IL36R “antigen-binding protein” is a single polypeptide (e.g.,an ScFv (single chain variable fragment)) or complex of more than onepolypeptide (e.g., a tetrameric IgG antibody) that binds specifically tothe IL36 receptor at the IL1RL2 subunit (IL-1Rrp2). IL-36R, in thecontext of binding of an antigen-binding protein thereto, refers toIL-1RL2. In an embodiment of the invention, the antigen-binding proteinis an antibody or antigen-binding fragment whether monospecific ormultispecific (e.g., bispecific) or monovalent or multivalent (e.g.,bivalent). A monovalent antigen-binding protein has a singleantigen-binding domain whereas a bivalent antigen-binding protein hastwo antigen-binding domains.

A polynucleotide includes DNA and RNA. The present invention includesany polynucleotide of the present invention which is operably linked toa promoter or other expression control sequence.

In general, a “promoter” or “promoter sequence” is a DNA regulatoryregion capable of binding an RNA polymerase in a cell (e.g., directly orthrough other promoter-bound proteins or substances) and initiatingtranscription of a coding sequence. A promoter sequence is, in general,bounded at its 3′ terminus by the transcription initiation site andextends upstream (5′ direction) to include the minimum number of basesor elements necessary to initiate transcription at any level. Within thepromoter sequence may be found a transcription initiation site(conveniently defined, for example, by mapping with nuclease S1), aswell as protein binding domains (consensus sequences) responsible forthe binding of RNA polymerase. The promoter may be operably associatedwith other expression control sequences, including enhancer andrepressor sequences or with a nucleic acid of the invention. Promoterswhich may be used to control gene expression include, but are notlimited to, cytomegalovirus (CMV) promoter (U.S. Pat. Nos. 5,385,839 and5,168,062), the SV40 early promoter region (Benoist, et al., (1981)Nature 290:304-310), the promoter contained in the 3′ long terminalrepeat of Rous sarcoma virus (Yamamoto, et al., (1980) Cell 22:787-797),the herpes thymidine kinase promoter (Wagner, et al., (1981) Proc. Natl.Acad. Sci. USA 78:1441-1445), the regulatory sequences of themetallothionein gene (Brinster, et al., (1982) Nature 296:39-42);prokaryotic expression vectors such as the beta-lactamase promoter(VIlla-Komaroff, et al., (1978) Proc. Natl. Acad. Sci. USA75:3727-3731), or the tac promoter (DeBoer, et al., (1983) Proc. Natl.Acad. Sci. USA 80:21-25); see also “Useful proteins from recombinantbacteria” in Scientific American (1980) 242:74-94; and promoter elementsfrom yeast or other fungi such as the Gal 4 promoter, the ADC (alcoholdehydrogenase) promoter, PGK (phosphoglycerol kinase) promoter or thealkaline phosphatase promoter.

A polynucleotide encoding a polypeptide is “operably linked” to apromoter or other expression control sequence when, in a cell, thesequence directs RNA polymerase mediated transcription of the codingsequence into RNA, preferably mRNA, which then may be RNA spliced (if itcontains introns) and, optionally, translated into a protein encoded bythe coding sequence.

Interleukin-36 Receptor (IL36R)

IL-36R is a member of the IL-1 receptor family that contains sixreceptor proteins that form four signaling complexes: IL-1RI, IL-18R,IL-33R, and IL-36R, and two decoy receptors and two negative regulators.IL-36R is a heterodimer that consists of a receptor subunit namedIL-1Rrp2 (also known as IL-1RL2, Interleukin 1 receptor-like 2 orInterleukin 1 receptor-related protein 2) and a co-receptor subunitInterleukin-1 receptor accessory protein, IL-1 RAcP. The receptor canrecognize three different agonists, IL-36α, IL-36β, and IL-36γ (alsoknown as IL-1F6, IL-1F8, and IL-1F9), to induce the expression ofinflammatory cytokines. There are also two receptor antagonists, IL-36Raand IL-38, which bind to IL-36 receptor and decrease the expression ofinflammatory cytokines. IL-36α, IL-36β, and IL-36γ signal through theIL-36R/IL-1RAcP receptor to activate NF-κB and MAPKs, such as p38 andJNK, and promote inflammatory responses.

In an embodiment of the invention, the Homo sapiens IL1RL2 sequence isavailable under Genbank accession number NP_003845.2. In an embodimentof the invention, the amino acid sequence of Homo sapiens IL1RL2 is setforth in SEQ ID NO: 177.

In an embodiment of the invention, Homo sapiens IL-1RAcP sequence isavailable under Genbank accession no. NP_002173.1. In an embodiment ofthe invention, the amino acid sequence of Homo sapiens IL-1RAcP is setforth in SEQ ID NO: 178.

Anti-L36 Antibodies and Antigen-Binding Fragments Thereof

The present invention provides antigen-binding proteins, such asantibodies (e.g., human antibodies) and antigen-binding fragmentsthereof, that specifically bind to IL36R protein or an antigenicfragment thereof. Antigen-binding proteins that bind to the same epitopeon IL36R as, or compete for binding to IL36R with any of theantigen-binding proteins set forth herein are also part of the presentinvention.

The term “antibody”, as used herein, refers to immunoglobulin moleculescomprising four polypeptide chains, two heavy chains (HCs) and two lightchains (LCs) inter-connected by disulfide bonds (i.e., “full antibodymolecules”), as well as multimers thereof—for example H4H14699P2;H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P;H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2. In anembodiment of the invention, each heavy chain (HC) comprises a heavychain variable region (“HCVR” or “V_(H)”) (e.g., SEQ ID NO: 2, 18, 34,50, 66, 82, 98, 114, 130, 138, 154 or 170 or a variant thereof) and aheavy chain constant region (including domains CH1, CH2 and CH3); andeach light chain (LC) comprises a light chain variable region (“LCVR or“V_(L)”) (e.g., SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146 or 162or a variant thereof) and a light chain constant region (C_(L)). TheV_(H) and V_(L) regions can be further subdivided into regions ofhypervariability, termed complementarity determining regions (CDR),interspersed with regions that are more conserved, termed frameworkregions (FR). Each V_(H) and V_(L) comprises three CDRs and four FRs,arranged from amino-terminus to carboxy-terminus in the following order:FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments of theinvention, the FRs of the antibody (or antigen binding fragment thereof)are identical to the human germline sequences, or are naturally orartificially modified.

Typically, the variable domains of both the heavy and lightimmunoglobulin chains comprise three hypervariable regions, also calledcomplementarity determining regions (CDRs), located within relativelyconserved framework regions (FR). In general, from N-terminal toC-terminal, both light and heavy chains variable domains comprise FR1,CDR1, FR2, CDR2, FR3, CDR3 and FR4. In an embodiment of the invention,the assignment of amino acids to each domain is in accordance with thedefinitions of Sequences of Proteins of Immunological Interest, Kabat,et al., National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ.No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat, etal., (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al., (1987) J Mol.Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883.

For example, the present invention provides an antigen-binding proteinthat includes (a) a heavy chain immunoglobulin comprising the CDR-H1,CDR-H2 and CDR-H3 of a heavy chain immunoglobulin comprising an aminoacid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114,130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216,220 or 224 and at least 70, 80 or 90% amino acid sequence identity tothe amino acid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82,98, 114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208,212, 216, 220 or 224; and (b) an light chain immunoglobulin comprisingthe CDR-L1, CDR-L2 and CDR-L3 of a light chain immunoglobulin comprisingan amino acid sequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90,106, 122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218,222 or 226 and at least 70, 80 or 90% amino acid sequence identity tothe amino acid sequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90,106, 122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218,222 or 226. In an embodiment of the invention, the antigen-bindingprotein includes (i) a heavy chain immunoglobulin that comprises CDR-H1,CDR-H2 and CDR-H3 of a heavy chain immunoglobulin that comprises theamino acid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98,114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212,216, 220 or 224; or a variant thereof; and (ii) a light chainimmunoglobulin that comprises CDR-L1, CDR-L2 and CDR-L3 of a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194,198, 202, 206, 210, 214, 218, 222 or 226; or a variant thereof.

In an embodiment of the invention, an antigen-binding protein of thepresent invention includes a heavy chain immunoglobulin that comprises aV_(H) including CDR-H1, CDR-H2 and CDR-H3, wherein the:

CDR-H1 comprises the amino acid sequence set forth in SEQ ID NO: 4, 20,36, 52, 68, 84, 100, 116, 132, 140, 156 or 172, or a variant thereof;CDR-H2 comprises the amino acid sequence set forth in SEQ ID NO: 6, 22,38, 54, 70, 86, 102, 118, 134, 142, 158 or 174, or a variant thereof;andCDR-H3 comprises the amino acid sequence set forth in SEQ ID NO: 8, 24,40, 56, 72, 88, 104, 120, 136, 144, 160 or 176, or a variant thereof;anda light chain immunoglobulin that comprises a V_(L) including CDR-L1,CDR-L2 and CDR-L3, wherein the:CDR-L1 comprises the amino acid sequence set forth in SEQ ID NO: 12, 28,44, 60, 76, 92, 108, 124, 124, 148 or 164, or a variant thereof;CDR-L2 comprises the amino acid sequence set forth in SEQ ID NO: 14, 30,46, 62, 78, 94, 110, 126, 126, 150 or 166, or a variant thereof; andCDR-L3 comprises the amino acid sequence set forth in SEQ ID NO: 16, 32,48, 64, 80, 96, 112, 128, 128, 152 or 168, or a variant thereof.

The present invention includes monoclonal anti-IL36R antigen-bindingproteins, e.g., antibodies and antigen-binding fragments thereof, aswell as monoclonal compositions comprising a plurality of isolatedmonoclonal antigen-binding proteins. The term “monoclonal antibody” or“mAb”, as used herein, refers to a member of a population ofsubstantially homogeneous antibodies, i.e., the antibody moleculescomprising the population are identical in amino acid sequence exceptfor possible naturally occurring mutations that may be present in minoramounts. A “plurality” of such monoclonal antibodies and fragments in acomposition refers to a concentration of identical (i.e., as discussedabove, in amino acid sequence except for possible naturally occurringmutations that may be present in minor amounts) antibodies and fragmentswhich is above that which would normally occur in nature, e.g., in theblood of a host organism such as a mouse or a human.

In an embodiment of the invention, an anti-IL36R antigen-bindingprotein, e.g., antibody or antigen-binding fragment comprises a heavychain constant domain, e.g., of the type IgA (e.g., IgA1 or IgA2), IgD,IgE, IgG (e.g., IgG1, IgG2, IgG3 and IgG4) or IgM. In an embodiment ofthe invention, an antigen-binding protein, e.g., antibody orantigen-binding fragment, comprises a light chain constant domain, e.g.,of the type kappa or lambda.

The present invention includes human antigen-binding proteins. The term“human” antigen-binding protein, such as an antibody or antigen-bindingfragment, as used herein, includes antibodies and fragments havingvariable and constant regions derived from human germline immunoglobulinsequences whether in a human cell or grafted into a non-human cell,e.g., a mouse cell. See e.g., U.S. Pat. No. 8,502,018, 6,596,541 or5,789,215. The human mAbs of the invention may include amino acidresidues not encoded by human germline immunoglobulin sequences (e.g.,mutations introduced by random or site-specific mutagenesis in vitro orby somatic mutation in vivo), for example in the CDRs and in particularCDR3. However, the term “human antibody”, as used herein, is notintended to include mAbs in which CDR sequences derived from thegermline of another mammalian species (e.g., mouse) have been graftedonto human FR sequences. The term includes antibodies recombinantlyproduced in a non-human mammal or in cells of a non-human mammal. Theterm is not intended to include natural antibodies directly isolatedfrom a human subject.

The present invention includes anti-IL36R chimeric antigen-bindingproteins, e.g., antibodies and antigen-binding fragments thereof, andmethods of use thereof. As used herein, a “chimeric antibody” is anantibody having the variable domain from a first antibody and theconstant domain from a second antibody, where the first and secondantibodies are from different species. (see e.g., U.S. Pat. No.4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855).

The term “recombinant” antigen-binding proteins, such as antibodies orantigen-binding fragments thereof, refers to such molecules created,expressed, isolated or obtained by technologies or methods known in theart as recombinant DNA technology which include, e.g., DNA splicing andtransgenic expression. The term includes antibodies expressed in anon-human mammal (including transgenic non-human mammals, e.g.,transgenic mice), or a cell (e.g., CHO cells) such as a cellularexpression system or isolated from a recombinant combinatorial humanantibody library.

Recombinant anti-IL36R antigen-binding proteins, e.g., antibodies andantigen-binding fragments, disclosed herein may also be produced in anE. coli/T7 expression system. In this embodiment, nucleic acids encodingthe anti-IL36R antibody immunoglobulin molecules of the invention (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2)may be inserted into a pET-based plasmid and expressed in the E. coli/T7system. For example, the present invention includes methods forexpressing an antibody or antigen-binding fragment thereof orimmunoglobulin chain thereof in a host cell (e.g., bacterial host cellsuch as E. coli such as BL21 or BL21DE3) comprising expressing T7 RNApolymerase in the cell which also includes a polynucleotide encoding animmunoglobulin chain that is operably linked to a T7 promoter. Forexample, in an embodiment of the invention, a bacterial host cell, suchas an E. coli, includes a polynucleotide encoding the T7 RNA polymerasegene operably linked to a lac promoter and expression of the polymeraseand the chain is induced by incubation of the host cell with IPTG(isopropyl-beta-D-thiogalactopyranoside). See U.S. Pat. Nos. 4,952,496and 5,693,489 or Studier & Moffatt, Use of bacteriophage T7 RNApolymerase to direct selective high-level expression of cloned genes, J.Mol. Biol. 1986 May 5; 189(1): 113-30.

There are several methods by which to produce recombinant antibodieswhich are known in the art. One example of a method for recombinantproduction of antibodies is disclosed in U.S. Pat. No. 4,816,567.

Transformation can be by any known method for introducingpolynucleotides into a host cell. Methods for introduction ofheterologous polynucleotides into mammalian cells are well known in theart and include dextran-mediated transfection, calcium phosphateprecipitation, polybrene-mediated transfection, protoplast fusion,electroporation, encapsulation of the polynucleotide(s) in liposomes,biolistic injection and direct microinjection of the DNA into nuclei. Inaddition, nucleic acid molecules may be introduced into mammalian cellsby viral vectors. Methods of transforming cells are well known in theart. See, for example, U.S. Pat. Nos. 4,399,216; 4,912,040; 4,740,461and 4,959,455. Thus, the present invention includes recombinant methodsfor making an anti-IL36R antigen-binding protein, such as an antibody orantigen-binding fragment thereof of the present invention, or animmunoglobulin chain thereof, comprising (i) introducing one or morepolynucleotides (e.g., including the nucleotide sequence in any one ormore of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27,29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63,65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99,101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125, 127,129, 131, 133, 135, 137, 139, 141, 143, 145, 147, 149, 151, 153, 155,157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 179, 181, 183, 185,187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213,215, 217, 219, 221, 223 and/or 225; or a variant thereof) encoding lightand/or heavy immunoglobulin chains of the antigen-binding protein, e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2,for example, wherein the polynucleotide is in a vector, and/orintegrated into a host cell chromosome and/or is operably linked to apromoter (ii) culturing the host cell (e.g., CHO or Pichia or Pichiapastoris) under conditions favorable to expression of the polynucleotideand, (iii) optionally, isolating the antigen-binding protein (e.g.,antibody or fragment) or chain from the host cell and/or medium in whichthe host cell is grown. When making an antigen-binding protein (e.g.,antibody or antigen-binding fragment) comprising more than oneimmunoglobulin chain, e.g., an antibody that comprises two heavyimmunoglobulin chains and two light immunoglobulin chains, co-expressionof the chains in a single host cell leads to association of the chains,e.g., in the cell or on the cell surface or outside the cell if suchchains are secreted, so as to form the antigen-binding protein (e.g.,antibody or antigen-binding fragment). The methods of the presentinvention include those wherein only a heavy immunoglobulin chain oronly a light immunoglobulin chain or both (e.g., any of those discussedherein including mature fragments and/or variable domains thereof) areexpressed in a cell. Such single chains are useful, for example, asintermediates in the expression of an antibody or antigen-bindingfragment that includes such a chain. For example, the present inventionalso includes anti-IL36R antigen-binding proteins, such as antibodiesand antigen-binding fragments thereof, comprising a heavy chainimmunoglobulin (or variable domain thereof or comprising the CDRsthereof) encoded by a polynucleotide comprising the nucleotide sequencesset forth in SEQ ID NO: 1, 17, 33, 49, 65, 81, 97, 113, 129, 137, 153,169, 179, 183, 187, 191, 195, 199, 203, 207, 211, 215, 219 or 223; and alight chain immunoglobulin (or variable domain thereof or comprising theCDRs thereof) encoded by the nucleotide sequence set forth in SEQ ID NO:9, 25, 41, 57, 73, 89, 105, 121, 145, 161, 181, 185, 189, 193, 197, 201,205, 209, 213, 217, 221 or 225 which are the product of such productionmethods, and, optionally, the purification methods set forth herein. Forexample, in an embodiment of the invention, the product of the method isan anti-IL36R antigen-binding protein which is an antibody or fragmentcomprising a heavy chain immunoglobulin or V_(H) comprising the aminoacid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114,130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216,220 or 224 and a light chain immunoglobulin or V_(L) comprising theamino acid sequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106,122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or226.

In an embodiment of the invention, a method for making an anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragmentthereof, includes a method of purifying the antigen-binding protein,e.g., by column chromatography, precipitation and/or filtration. Theproduct of such a method also forms part of the present invention.

Eukaryotic and prokaryotic host cells, including mammalian cells, may beused as hosts for expression of an anti-IL36R antigen-binding protein(e.g., antibody or antigen-binding fragment thereof). Such host cellsare well known in the art and many are available from the American TypeCulture Collection (ATCC). These host cells include, inter alia, Chinesehamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamsterkidney (BHK) cells, monkey kidney cells (COS), human hepatocellularcarcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells anda number of other cell lines. Mammalian host cells include human, mouse,rat, dog, monkey, pig, goat, bovine, horse and hamster cells. Other celllines that may be used are insect cell lines (e.g., Spodopterafrugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plantcells and fungal cells. Fungal cells include yeast and filamentousfungus cells including, for example, Pichia, Pichia pastoris, Pichiafinlandica, Pichia trehalophila, Pichia koclamae, Pichiamembranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindnen), Pichiaopuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum,Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichia sp.,Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha,Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillusnidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei,Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusariumvenenatum, Physcomitrella patens and Neurospora crassa. The presentinvention includes an isolated host cell (e.g., a CHO cell or any typeof host cell set forth above) comprising an antigen-binding protein,such as H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P orH4H14760P2; and/or a polynucleotide encoding one or more immunoglobulinchains thereof.

The present invention also includes a cell which is expressing IL36R oran antigenic fragment or fusion thereof (e.g., His₆, Fc and/or myc)which is bound by an antigen-binding protein of the present inventione.g., H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P orH4H14760P2, e.g., wherein the cell is in the body of a subject or is invitro.

In addition, the present invention also provides a complex comprising ananti-IL36R antigen-binding protein, e.g., antibody or antigen-bindingfragment, discussed herein complexed with IL36R polypeptide or anantigenic fragment thereof or fusion thereof and/or with a secondaryantibody or antigen-binding fragment thereof (e.g., detectably labeledsecondary antibody) that binds specifically to the anti-IL36R antibodyor fragment. In an embodiment of the invention, the complex is in vitro(e.g., is immobilized to a solid substrate) or is in the body of asubject.

The term “specifically binds” refers to those antigen-binding proteins(e.g., mAbs) having a binding affinity to an antigen, such as IL36Rprotein, expressed as Ko, of at least about 58 nM (e.g., 10⁻⁹ M, 10⁻¹⁰M,10⁻¹¹ M, or 10⁻¹² M), as measured by real-time, label free bio-layerinterferometry assay, for example, at 25° C. or 37° C., e.g., an Octet®HTX biosensor, or by surface plasmon resonance, e.g., BIACORE™, or bysolution-affinity ELISA. The present invention includes antigen-bindingproteins that specifically bind to IL36R protein. In an embodiment ofthe invention, an anti-IL36R antigen-binding protein comprises a K_(D)value, for binding to human and/or Macaca fascicularis IL36R, whichvalue is set forth in any of Tables 4-1 to 4-8.

The terms “antigen-binding portion” or “antigen-binding fragment” of anantibody or antigen-binding protein, and the like, as used herein,include any naturally occurring, enzymatically obtainable, synthetic, orgenetically engineered polypeptide or glycoprotein that specificallybinds an antigen to form a complex. Non-limiting examples ofantigen-binding fragments include: (i) Fab fragments; (ii) F(ab′)₂fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv(scFv) molecules; and (vi) dAb fragments; consisting of the amino acidresidues that mimic the hypervariable region of an antibody (e.g., anisolated complementarity determining region (CDR) such as a CDR3peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineeredmolecules, such as domain-specific antibodies, single domain antibodies,domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies,diabodies, triabodies, tetrabodies, minibodies and small modularimmunopharmaceuticals (SMIPs), are also encompassed within theexpression “antigen-binding fragment,” as used herein. In an embodimentof the invention, the antigen-binding fragment comprises three or moreCDRs of H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P orH4H14760P2 (e.g., CDR-H1, CDR-H2 and CDR-H3; or CDR-L1, CDR-L2 andCDR-L3).

An antigen-binding fragment of an antibody will, in an embodiment of theinvention, comprise at least one variable domain. The variable domainmay be of any size or amino acid composition and will generally compriseat least one CDR, which is adjacent to or in frame with one or moreframework sequences. In antigen-binding fragments having a V_(H) domainassociated with a V_(L) domain, the V_(H) and V_(L) domains may besituated relative to one another in any suitable arrangement. Forexample, the variable region may be dimeric and contain V_(H)-V_(H),V_(H)-V_(L) or V_(L)-V_(L) dimers. Alternatively, the antigen-bindingfragment of an antibody may contain a monomeric V_(H) or V_(L) domain.

In certain embodiments, an antigen-binding fragment of an antibody maycontain at least one variable domain covalently linked to at least oneconstant domain. Non-limiting, exemplary configurations of variable andconstant domains that may be found within an antigen-binding fragment ofan antibody of the present invention include: (i) V_(H)-C_(H)1; (ii)V_(H)-C_(H)2; (iii) V_(H)-C_(H)3; (iv) V_(H)-C_(H)1-C_(H)2; (v)V_(H)-C_(H)1-C_(H)2-C_(H)3; (vi) V_(H)-C_(H)2-C_(H)3; (vii) V_(H)-C_(L);(viii) V_(L)—C_(H)1; (ix) V_(L)-C_(H)2; (x) V_(L)-C_(H)3; (xi)V_(L)-C_(H)1-C_(H)2; (xii) V_(L)-C_(H)1-C_(H)2-C_(H)3; (xiii)V_(L)-C_(H)2-C_(H)3; and (xiv) V_(L)-C_(L). In any configuration ofvariable and constant domains, including any of the exemplaryconfigurations listed above, the variable and constant domains may beeither directly linked to one another or may be linked by a full orpartial hinge or linker region. A hinge region may consist of at least 2(e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, which result in aflexible or semi-flexible linkage between adjacent variable and/orconstant domains in a single polypeptide molecule. Moreover, anantigen-binding fragment of an antibody of the present invention maycomprise a homo-dimer or hetero-dimer (or other multimer) of any of thevariable and constant domain configurations listed above in non-covalentassociation with one another and/or with one or more monomeric V_(H) orV_(L) domain (e.g., by disulfide bond(s)).

Antigen-binding proteins (e.g., antibodies and antigen-bindingfragments) may be monospecific or multi-specific (e.g., bispecific).Multispecific antigen-binding proteins are discussed further herein.

In specific embodiments, antigen-binding proteins of the presentinvention (e.g., an antibody or antibody fragment) may be conjugated toa moiety such a ligand, a detectable label or a therapeutic moiety(“immunoconjugate”), a second anti-IL36R antibody, or any othertherapeutic moiety.

“Isolated” antigen-binding proteins (e.g., antibodies or antigen-bindingfragments thereof), polypeptides, polynucleotides and vectors, are atleast partially free of other biological molecules from the cells orcell culture from which they are produced. Such biological moleculesinclude nucleic acids, proteins, other antibodies or antigen-bindingfragments, lipids, carbohydrates, or other material such as cellulardebris and growth medium. An isolated antigen-binding protein mayfurther be at least partially free of expression system components suchas biological molecules from a host cell or of the growth mediumthereof. Generally, the term “isolated” is not intended to refer to acomplete absence of such biological molecules or to an absence of water,buffers, or salts or to components of a pharmaceutical formulation thatincludes the antigen-binding proteins (e.g., antibodies orantigen-binding fragments).

The present invention includes antigen-binding proteins, e.g.,antibodies or antigen-binding fragments, that bind to the same epitopeas an antigen-binding protein of the present invention (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2).

The term “epitope” refers to an antigenic determinant (e.g., on IL1RL2)that interacts with a specific antigen-binding site of anantigen-binding protein, e.g., a variable region of an antibodymolecule, known as a paratope. A single antigen may have more than oneepitope. Thus, different antibodies may bind to different areas on anantigen and may have different biological effects. The term “epitope”may also refer to a site on an antigen to which B and/or T cells respondand/or to a region of an antigen that is bound by an antibody. Epitopesmay be defined as structural or functional. Functional epitopes aregenerally a subset of the structural epitopes and have those residuesthat directly contribute to the affinity of the interaction. Epitopesmay be linear or conformational, that is, composed of non-linear aminoacids. In certain embodiments, epitopes may include determinants thatare chemically active surface groupings of molecules such as aminoacids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, incertain embodiments, may have specific three-dimensional structuralcharacteristics, and/or specific charge characteristics.

Methods for determining the epitope of an antigen-binding protein, e.g.,antibody or fragment or polypeptide, include alanine scanning mutationalanalysis, peptide blot analysis (Reineke (2004) Methods Mol. Biol. 248:443-63), peptide cleavage analysis, crystallographic studies and NMRanalysis. In addition, methods such as epitope excision, epitopeextraction and chemical modification of antigens can be employed (Tomer(2000) Prot. Sci. 9: 487-496). Another method that can be used toidentify the amino acids within a polypeptide with which anantigen-binding protein (e.g., antibody or fragment or polypeptide)interacts is hydrogen/deuterium exchange detected by mass spectrometry.See, e.g., Ehring (1999) Analytical Biochemistry 267: 252-259; Engen andSmith (2001) Anal. Chem. 73: 256A-265A.

The present invention includes antigen-binding proteins that compete forbinding to IL36R, e.g., a variant IL36R epitope as discussed herein,with an antigen-binding protein of the present invention, e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2.The term “competes” as used herein, refers to an antigen-binding protein(e.g., antibody or antigen-binding fragment thereof) that binds to anantigen (e.g., IL1RL2) and inhibits or blocks the binding of anotherantigen-binding protein (e.g., antibody or antigen-binding fragmentthereof) to the antigen. The term also includes competition between twoantigen-binding proteins e.g., antibodies, in both orientations, i.e., afirst antibody that binds and blocks binding of second antibody and viceversa. In certain embodiments, the first antigen-binding protein (e.g.,antibody) and second antigen-binding protein (e.g., antibody) may bindto the same epitope. Alternatively, the first and second antigen-bindingproteins (e.g., antibodies) may bind to different, but, for example,overlapping epitopes, wherein binding of one inhibits or blocks thebinding of the second antibody, e.g., via steric hindrance. Competitionbetween antigen-binding proteins (e.g., antibodies) may be measured bymethods known in the art, for example, by a real-time, label-freebio-layer interferometry assay. Also, binding competition betweenanti-IL36R antigen-binding proteins (e.g., monoclonal antibodies (mAbs))can be determined using a real time, label-free bio-layer interferometryassay on an Octet RED384 biosensor (Pall ForteBio Corp.).

Typically, an antibody or antigen-binding fragment of the inventionwhich is modified in some way retains the ability to specifically bindto IL36R, e.g., retains at least 10% of its IL36R binding activity (whencompared to the parental antibody) when that activity is expressed on amolar basis. Preferably, an antibody or antigen-binding fragment of theinvention retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or moreof the IL36R binding affinity as the parental antibody. It is alsointended that an antibody or antigen-binding fragment of the inventionmay include conservative or non-conservative amino acid substitutions(referred to as “conservative variants” or “function conserved variants”of the antibody) that do not substantially alter its biologic activity.

A “variant” of a polypeptide, such as an immunoglobulin chain (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2V_(H), V_(L), HC or LC), refers to a polypeptide comprising an aminoacid sequence that is at least about 70-99.9% (e.g., 70, 72, 74, 75, 76,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 99.5, 99.9%) identical or similar to a referenced amino acidsequence that is set forth herein (e.g., any of SEQ ID NOs: 2, 4, 6, 8,10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112,114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,170, 172, 174, 176, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198,200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224 or 226);when the comparison is performed by a BLAST algorithm wherein theparameters of the algorithm are selected to give the largest matchbetween the respective sequences over the entire length of therespective reference sequences (e.g., expect threshold: 10; word size:3; max matches in a query range: 0; BLOSUM 62 matrix; gap costs:existence 11, extension 1; conditional compositional score matrixadjustment).

A “variant” of a polynucleotide refers to a polynucleotide comprising anucleotide sequence that is at least about 70-99.9% (e.g., 70, 72, 74,75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 99.5, 99.9%) identical to a referenced nucleotidesequence that is set forth herein (e.g., any of SEQ ID NOs: 1, 3, 5, 7,9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43,45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79,81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111,113, 115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139,141, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167,169, 171, 173, 175, 179, 183, 187, 191, 195, 199, 203, 207, 211, 215,219, 223, 181, 185, 189, 193, 197, 201, 205, 209, 213, 217, 221 and/or225); when the comparison is performed by a BLAST algorithm wherein theparameters of the algorithm are selected to give the largest matchbetween the respective sequences over the entire length of therespective reference sequences (e.g., expect threshold: 10; word size:28; max matches in a query range: 0; match/mismatch scores: 1, −2; gapcosts: linear).

The following references relate to BLAST algorithms often used forsequence analysis: BLAST ALGORITHMS: Altschul et al. (2005) FEBS J.272(20): 5101-5109; Altschul, S. F., et al., (1990) J. Mol. Biol.215:403-410; Gish, W., et al., (1993) Nature Genet. 3:266-272; Madden,T. L., et al., (1996) Meth. Enzymol. 266:131-141; Altschul, S. F., etal., (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J., et al., (1997)Genome Res. 7:649-656; Wootton, J. C., et al., (1993) Comput. Chem.17:149-163; Hancock, J. M. et al., (1994) Comput. Appl. Biosci.10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, M. O., et al., “A model ofevolutionary change in proteins.” in Atlas of Protein Sequence andStructure, (1978) vol. 5, suppl. 3. M. O. Dayhoff (ed.), pp. 345-352,Natl. Biomed. Res. Found., Washington, D.C.; Schwartz, R. M., et al.,“Matrices for detecting distant relationships.” in Atlas of ProteinSequence and Structure, (1978) vol. 5, suppl. 3.” M. O. Dayhoff (ed.),pp. 353-358, Natl. Biomed. Res. Found., Washington, D.C.; Altschul, S.F., (1991) J. Mol. Biol. 219:555-565; States, D. J., et al., (1991)Methods 3:66-70; Henikoff, S., et al., (1992) Proc. Natl. Acad. Sci. USA89:10915-10919; Altschul, S. F., et al., (1993) J. Mol. Evol.36:290-300; ALIGNMENT STATISTICS: Karlin, S., et al., (1990) Proc. Natl.Acad. Sci. USA 87:2264-2268; Karlin, S., et al., (1993) Proc. Natl.Acad. Sci. USA 90:5873-5877; Dembo, A., et al., (1994) Ann. Prob.22:2022-2039; and Altschul, S. F. “Evaluating the statisticalsignificance of multiple distinct local alignments.” in Theoretical andComputational Methods in Genome Research (S. Suhai, ed.), (1997) pp.1-14, Plenum, N.Y.

Anti-IL36R antigen-binding proteins, e.g., antibodies andantigen-binding fragments thereof of the present invention, in anembodiment of the invention, include a heavy chain immunoglobulin orvariable region thereof having at least 70% (e.g., 80%, 85%, 90%, 95%,99%) amino acid sequence identity to the amino acids set forth in SEQ IDNO: 2, 18, 34, 50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184, 188,192, 196, 200, 204, 208, 212, 216, 220 or 224; and/or a light chainimmunoglobulin or variable region thereof having at least 70% (e.g.,80%, 85%, 90%, 95%, 99%) amino acid sequence identity to the amino acidsset forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182,186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or 226.

In addition, an anti-IL36R antigen-binding protein may include apolypeptide comprising an amino acid sequence that is set forth hereinexcept for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) mutationssuch as, for example, missense mutations (e.g., conservativesubstitutions), non-sense mutations, deletions, or insertions. Forexample, the present invention includes anti-IL36R antigen-bindingproteins which include an immunoglobulin light chain (or V_(L)) variantcomprising the amino acid sequence set forth in SEQ ID NO: 10, 26, 42,58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210,214, 218, 222 or 226 but having one or more of such mutations and/or animmunoglobulin heavy chain (or V_(H)) variant comprising the amino acidsequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130,138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or224 but having one or more of such mutations. In an embodiment of theinvention, an anti-IL36R antigen-binding protein includes animmunoglobulin light chain variant comprising CDR-L1, CDR-L2 and CDR-L3wherein one or more (e.g., 1 or 2 or 3) of such CDRs has one or more ofsuch mutations (e.g., conservative substitutions) and/or animmunoglobulin heavy chain variant comprising CDR-H1, CDR-H2 and CDR-H3wherein one or more (e.g., 1 or 2 or 3) of such CDRs has one or more ofsuch mutations (e.g., conservative substitutions).

Embodiments of the present invention also include antigen-bindingproteins, e.g., anti-IL36R antibodies and antigen-binding fragmentsthereof, that comprise immunoglobulin V_(H)S and V_(L)S; or HCs and LCs,which comprise a variant amino acid sequence having 70% or more (e.g.,80%, 85%, 90%, 95%, 97% or 99%) overall amino acid sequence identity orsimilarity to the amino acid sequences of the corresponding V_(H)S,V_(L)S, HCs or LCs specifically set forth herein, but wherein theCDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of suchimmunoglobulins are not variants and comprise the amino acid sequencesspecifically set forth herein. Thus, in such embodiments, the CDRswithin variant antigen-binding proteins are not, themselves, variants.

A “conservatively modified variant” or a “conservative substitution”,e.g., of an immunoglobulin chain set forth herein, refers to a variantwherein there is one or more substitutions of amino acids in apolypeptide with other amino acids having similar characteristics (e.g.charge, side-chain size, hydrophobicity/hydrophilicity, backboneconformation and rigidity, etc.). Such changes can frequently be madewithout significantly disrupting the biological activity of the antibodyor fragment. Those of skill in this art recognize that, in general,single amino acid substitutions in non-essential regions of apolypeptide do not substantially alter biological activity (see, e.g.,Watson et al. (1987) Molecular Biology of the Gene, TheBenjamin/Cummings Pub. Co., p. 224 (4^(th) Ed.)). In addition,substitutions of structurally or functionally similar amino acids areless likely to significantly disrupt biological activity. The presentinvention includes anti-IL36R antigen-binding proteins comprising suchconservatively modified variant immunoglobulin chains.

Examples of groups of amino acids that have side chains with similarchemical properties include 1) aliphatic side chains: glycine, alanine,valine, leucine and isoleucine; 2) aliphatic-hydroxyl side chains:serine and threonine; 3) amide-containing side chains: asparagine andglutamine; 4) aromatic side chains: phenylalanine, tyrosine, andtryptophan; 5) basic side chains: lysine, arginine, and histidine; 6)acidic side chains: aspartate and glutamate, and 7) sulfur-containingside chains: cysteine and methionine. Preferred conservative amino acidssubstitution groups are: valine-leucine-isoleucine,phenylalanine-tyrosine, lysine-arginine, alanine-valine,glutamate-aspartate, and asparagine-glutamine. Alternatively, aconservative replacement is any change having a positive value in thePAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science256: 1443 45.

Anti-IL36R antigen-binding proteins set forth herein, e.g., comprisingvariant immunoglobulin chains, may exhibit one or more of the followingproperties:

-   -   Binds to human IL36R (e.g., IL1RL2) (e.g., fused to a        myc-myc-His₆ tag) with a K_(D) of about 2.18 nM to about 13.9        nM, e.g., at 25° C. or with a K_(D) of about 4.25 nM to about        29.5 nM, e.g., at 37° C.;    -   Binds to Macaca fascicularis IL36R (e.g., IL1RL2) (e.g., fused        to a myc-myc-Hise tag) with a K_(D) of about 7.87 nM to about        34.4 nM, e.g., at 25° C. or with a K_(D) of about 14.4 nM to        about 58.2 nM, e.g., at 37° C.;    -   Binds to human IL36R (e.g., IL1RL2) (e.g., fused to a mouse        IgG2a) with a K_(D) of about 173 pM to about 5.79 nM, e.g., at        25° C. or with a K_(D) of about 205 pM to about 28.7 nM, e.g.,        at 37° C.;    -   Binds to human IL36R (e.g., IL1RL2) (e.g., fused to IL1RAcP        extracellular domain expressed with mouse IgG2a Fc tag) with a        K_(D) of about 212 pM to about 14 nM, e.g., at 25° C. or with a        K_(D) of about 264 pM to about 40.9 nM, e.g., at 37° C.;    -   Competes with one or more of the following anti-IL36R        antibodies: H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;        H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2;        H4H14757P; H4H14758P and/or H4H14760P2 for binding to IL36R        (e.g., IL1RL2), optionally with the proviso that such anti-IL36R        antibody or fragment which competes with H4H14699P2; H4H14700P2;        H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P;        H4H14732P2; H4H14734P2; H4H14757P; H4H14758P and/or H4H14760P2        is not antibody APE3798; APE4086; APE5125/APE5100; APE5216;        APE5281; APE5214/APE4881; APE5280; APE5257; APE5258/APE5076;        APE5212; APE5213/5066; APE5211; APE5217/APE5060; APE3849;        APE3850; APE5600; APE5598; APE5627; APE6064; APE6060; APE6157;        APE6155/APE6917; APE6194; APE3847; APE5713; APE6083;        APE6903/APE7247; APE6904; and/or APE6907 (e.g., APE6155) or an        antigen-binding fragment thereof or antigen-binding protein        comprising the CDRs or variable regions thereof (see        WO2016/168542);    -   Blocks activation of NFκB by IL-36R (e.g., human or Macaca        fascicularis) in the presence of IL-1RAcP and ligand such as        hIL-36α, hIL-36β, and/or hIL-36γ, e.g., wherein the NFκB is in a        host cell, such as HEK293, e.g., containing NFκB response        element (5×)-luciferase-IRES-GFP, e.g., with an IC₅₀ of about        1×10⁻¹⁰ M⁻⁷×10⁻⁹ M;    -   Prevents or ameliorates skin inflammation (e.g., chronic or        acute) or reduces skin thickness or total pathology score or        reduces pro-inflammatory cytokine levels (e.g., KC-GRO, IL6,        IL1b and/or TNFalpha) in a subject suffering from skin        inflammation (e.g., chronic or acute), e.g., chemically-induced        skin inflammation (e.g., imiquimod-induced), e.g., in a mouse        such as a mouse displaying symptoms of human DITRA (Deficiency        of Interleukin Thirty-six Receptor Antagonist) disease, which        disease is described in e.g., Marrakchi et al.,        Interleukin-36-receptor antagonist deficiency and generalized        pustular psoriasis, N Engl J Med 365:620-628 (2011)—e.g.,        relative to a subject not treated with such an antigen-binding        protein; and/or    -   Prevents or ameliorates colitis or colon inflammation, e.g.,        chemically-induced colitis or colon inflammation, e.g., induced        by dextran sulfate sodium (DSS) or oxazolone, or reduces fecal        levels of LCN2 polypeptide in a subject with such colitis or        inflammation e.g., in a mouse such as a DITRA mouse—e.g.,        relative to a subject not treated with such an antigen-binding        protein.    -   Protects residues (a) 113-119, 113-122, 116-119 and/or 116-122;        and/or (b) 264-271, 267-271, 268-271, 268-276, 268-277 and/or        271-276, of an IL36R polypeptide comprising the amino acid        sequence set forth in SEQ ID NO: 227, when bound, from digestion        with pepsin and/or Protease XIII (e.g., from Aspergillus saitoi)        and/or deuteration in the presence of deuterium (e.g., D₂O);*    -   Binds to IL36R, for example, the IL1RL2 subunit thereof, at        residues 113-119, 113-122, 116-119, 116-122, 264-271, 267-271,        268-271, 268-276, 268-277 and/or 271-276 of an IL36R polypeptide        comprising the amino acid sequence set forth in SEQ ID NO: 227;*    -   Binds Domain II of IL36R, e.g., with a coverage of about 80.0,        80.1, 81.0 or 81.5%;    -   Binds a polypeptide comprising the amino acid sequence        YKQILHLGKD (SEQ ID NO: 229) (amino acids 113-122 of SEQ ID        NO: 227) and/or GVETHVSFREHNYL (SEQ ID NO: 230) (amino acids        264-277 of SEQ ID NO: 227)    -   Inhibits IL36a, IL36p and/or IL36γ (e.g., at a concentration of        about 10 nM), e.g., in in vitro epidermal keratinocytes,        intestinal myofibroblasts and/or CD14+ monocytes, with an IC₅₀        of about 1, 2, 3, 4, 5 or 6 nM or 1-6 nM; and/or    -   Competitively inhibits IL36α, IL36β and/or IL36γ-mediated        activation of NFκB (e.g., an NFκB response element        (5×)-luciferase-IRES-GFP reporter in a cell such as HEK293) by        IL36R: for example, as measured in a Schild Assay format.        * Includes antigen-binding proteins that bind to a native IL36R        (IL1-RL2), e.g., as set forth under UniProt Accession No.        Q9HB29, at residues corresponding to those set forth in the        tagged IL36R polypeptide comprising the amino acid sequence set        forth in SEQ ID NO: 227. See below:

113-119: (SEQ ID NO: 228)YKQILHL (amino acids 113-119 of SEQ ID NO: 227); 113-122:(SEQ ID NO: 229) YKQILHLGKD (amino acids 113-122 of SEQ ID NO: 227);116-119: (SEQ ID NO: 231) ILHL (amino acids 116-119 of SEQ ID NO: 227);116-122: (SEQ ID NO: 232)ILHLGKD (amino acids 116-122 of SEQ ID NO: 227); 264-271:(SEQ ID NO: 233) GVETHVSF (amino acids 264-271 of SEQ ID NO: 227);267-271: (SEQ ID NO: 234) THVSF (amino acids 267-271 of SEQ ID NO: 227);268-271: (SEQ ID NO: 235) HVSF (amino acids 268-271 of SEQ ID NO: 227);268-276: (SEQ ID NO: 236)HVSFREHNL (amino acids 268-276 of SEQ ID NO: 227); 268-277:(SEQ ID NO: 237) HVSFREHNLY (amino acids 268-277 of SEQ ID NO: 227);271-276: (SEQ ID NO: 238)FREHNL (amino acids 271-276 of SEQ ID NO: 227).

See residues highlighted below in human IL36R (IL1RL2):

(SEQ ID NO: 117) MTGLVSLSYF PLSTRSCALQ SCRQPGLGMWSLLLCGLSIA LPLSVTADGC KDIFMKNEIL SASQPFAFNC TFPPITSGEV SVTWYKNSSKIPVSKIIQSR IHQDETWILF LPMEWGDSGV YQCVIKGRDS CHRIHVNLTV FEKHWCDTSIGGLPNLSDEY KQILHLGKDD SLTCHLHFPK SCVLGPIKWY KDCNEIKGER FTVLETRLLVSNVSAEDRGN YACQAILTHS GKQYEVLNGI TVSITERAGY GGSVPKIIYP KNHSIEVQLGTTLIVDCNVT DTKDNTNLRC WRVNNTLVDD YYDESKRIRE GVETHVSFRE HNLYTVNITFLEVKMEDYGL PFMCHAGVST AYTILQLPAP DFRAYLIGGL IALVAVAVSV VYIYNIFKIDIVLWYRSAFH STETIVDGKL YDAYVLYPKP HKESQRHAVD ALVLNILPEV LERQCGYKLFIFGRDEFPGQ AVANVIDENV KLCRRLIVIV VPESLGFGLL KNLSEEQIAV YSALIQDGMKVILIELEKIE DYTVMPESIQ YIKQKHGAIR WHGDFTEQSQ CMKTKFWKTV RYHMPPRPCRPFPPVQLLQH TPCYRTAGPE LGSRRKKCTLTTG

The present invention includes “neutralizing” or “antagonist” anti-IL36Rantigen-binding proteins, e.g., antibody or antigen-binding fragment,which includes molecules that inhibit an activity of IL36R to anydetectable degree (e.g., IL36 ligand binding).

“H4H14699P2”, “H4H14700P2”, “H4H14706P2”, “H4H14708P2”, “H4H14709P”,“H4H14728P”, “H4H14731P”, “H4H14732P2”, “H4H14734P2”, “H4H14757P”,“H4H14758P” and “H4H14760P2” refer to antigen-binding proteins, e.g.,antibodies and antigen-binding fragments thereof (includingmultispecific antigen-binding proteins), comprising the immunoglobulinheavy chain or variable region thereof (V_(H)) of SEQ ID NO: 2, 18, 34,50, 66, 82, 98, 114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200,204, 208, 212, 216, 220 or 224 (or a variant thereof), respectively; andthe immunoglobulin light chain or variable region thereof (V_(L)) of 10,26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202,206, 210, 214, 218, 222 or 226 (or a variant thereof), respectively; orthat comprise a heavy chain or V_(H) that comprises the CDRs thereof(CDR-H1 (or a variant thereof), CDR-H2 (or a variant thereof) and CDR-H3(or a variant thereof)) and/or a light chain or V_(L) that comprises theCDRs thereof (CDR-L1 (or a variant thereof), CDR-L2 (or a variantthereof) and CDR-L3 (or a variant thereof)), e.g., wherein theimmunoglobulin chains, variable regions and/or CDRs comprise thespecific amino acid sequences described below. In an embodiment of theinvention, the V_(H) is linked to an IgG constant heavy chain domain(e.g., IgG1 or IgG4) and/or the V_(L) is linked to a lambda or kappaconstant light chain domain.

Antibodies and antigen-binding fragments of the present inventioncomprise immunoglobulin chains including the amino acid sequences setforth herein as well as cellular and in vitro post-translationalmodifications to the antibody or fragment. For example, the presentinvention includes antibodies and antigen-binding fragments thereof thatspecifically bind to IL36R comprising heavy and/or light chain aminoacid sequences set forth herein (e.g., CDR-H1, CDR-H2, CDR-H3, CDR-L1,CDR-L2 and/or CDR-L3) as well as antibodies and fragments wherein one ormore asparagine, serine and/or threonine residues is glycosylated, oneor more asparagine residues is deamidated, one or more residues (e.g.,Met, Trp and/or His) is oxidized, the N-terminal glutamine ispyroglutamate (pyroE) and/or the C-terminal lysine is missing.

The present invention provides a vessel (e.g., a plastic or glass vial,e.g., with a cap or a chromatography column, hollow bore needle or asyringe cylinder) comprising an anti-IL36R antigen-binding protein ofthe present invention, e.g., H4H14699P2; H4H14700P2; H4H14706P2;H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2;H4H14757P; H4H14758P or H4H14760P2.

The present invention also provides an injection device comprising oneor more antigen-binding proteins (e.g., antibody or antigen-bindingfragment) that bind specifically to IL36R, e.g., H4H14699P2; H4H14700P2;H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2;H4H14734P2; H4H14757P; H4H14758P or H4H14760P2, or a pharmaceuticalcomposition thereof. The injection device may be packaged into a kit. Aninjection device is a device that introduces a substance into the bodyof a subject via a parenteral route, e.g., intramuscular, subcutaneousor intravenous. For example, an injection device may be a syringe (e.g.,pre-filled with the pharmaceutical composition, such as anauto-injector) which, for example, includes a cylinder or barrel forholding fluid to be injected (e.g., comprising the antibody or fragmentor a pharmaceutical composition thereof), a needle for piecing skinand/or blood vessels for injection of the fluid; and a plunger forpushing the fluid out of the cylinder and through the needle bore.

The present invention further provides methods for administering ananti-IL36R antigen-binding protein of the present invention, e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2,to a subject, comprising introducing the antigen-binding protein intothe body of the subject (e.g., a human), for example, parenterally. Forexample, the method comprises piercing the body of the subject with aneedle of a syringe and injecting the antigen-binding protein into thebody of the subject, e.g., into the vein, artery, tumor, muscular tissueor subcutis of the subject.

Preparation of Human Antibodies

Methods for generating human antibodies in transgenic mice are known inthe art. Any such known methods can be used in the context of thepresent invention to make human antibodies that specifically bind toIL36R (e.g., IL1RL2). In certain embodiments of the invention, theantibodies of the invention are obtained from mice immunized with IL36R(e.g., IL1RL2 polypeptide or an immunogenic fragment thereof), or with alive attenuated or inactivated virus, or with DNA encoding the proteinor fragment thereof. Alternatively, IL36R may be produced using standardbiochemical techniques and modified and used as immunogen. In certainembodiments of the invention, the immunogen may be an IL36R (e.g.,IL1RL2) polypeptide vaccine. In certain embodiments, one or more boosterinjections may be administered. In certain embodiments, the immunogenmay be a recombinant IL36R polypeptide (e.g., IL1RL2) expressed in E.coli or in any other eukaryotic or mammalian cells such as Chinesehamster ovary (CHO) cells.

Using VELOCIMMUNE® technology (see, for example, U.S. Pat. No.6,596,541, Regeneron Pharmaceuticals, VELOCIMMUNE®) or any other knownmethod for generating monoclonal antibodies, high affinity chimericantibodies to IL36R can be initially isolated having a human variableregion and a mouse constant region. The VELOCIMMUNE® technology involvesgeneration of a transgenic mouse having a genome comprising human heavyand light chain variable regions operably linked to endogenous mouseconstant region loci such that the mouse produces an antibody comprisinga human variable region and a mouse constant region in response toantigenic stimulation. The DNA encoding the variable regions of theheavy and light chains of the antibody are isolated and operably linkedto DNA encoding the human heavy and light chain constant regions. TheDNA is then expressed in a cell capable of expressing the fully humanantibody.

Generally, a VELOCIMMUNE® mouse is challenged with the antigen ofinterest, and lymphatic cells (such as B-cells) are recovered from themice that express antibodies. The lymphatic cells may be fused with amyeloma cell line to prepare immortal hybridoma cell lines, and suchhybridoma cell lines are screened and selected to identify hybridomacell lines that produce antibodies specific to the antigen of interest.DNA encoding the variable regions of the heavy chain and light chain maybe isolated and linked to desirable isotypic constant regions of theheavy chain and light chain. Such an antibody protein may be produced ina cell, such as a CHO cell. Alternatively, DNA encoding theantigen-specific chimeric antibodies or the variable domains of thelight and heavy chains may be isolated directly from antigen-specificlymphocytes.

Initially, high affinity chimeric antibodies are isolated having a humanvariable region and a mouse constant region. The antibodies arecharacterized and selected for desirable characteristics, includingaffinity, selectivity, epitope, etc. The mouse constant regions arereplaced with a desired human constant region to generate the fullyhuman antibody of the invention, for example wild-type or modified IgG1or IgG4. While the constant region selected may vary according tospecific use, high affinity antigen-binding and target specificitycharacteristics reside in the variable region.

Anti-IL36R Antibodies Comprising Fc Variants

According to certain embodiments of the present invention, anti-IL36Rantigen-binding proteins, e.g., antibodies or antigen-binding fragments,are provided comprising an Fc domain comprising one or more mutations,which, for example, enhance or diminish antibody binding to the FcRnreceptor, e.g., at acidic pH as compared to neutral pH. For example, thepresent invention includes anti-IL36R antibodies comprising a mutationin the C_(H)2 or a C_(H)3 region of the Fc domain, wherein themutation(s) increases the affinity of the Fc domain to FcRn in an acidicenvironment (e.g., in an endosome where pH ranges from about 5.5 toabout 6.0). Such mutations may result in an increase in serum half-lifeof the antibody when administered to an animal. Non-limiting examples ofsuch Fc modifications include, e.g., a modification at position 250(e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., L/Y/F/W or T),254 (e.g., S or T), and 256 (e.g., S/R/Q/E/D or T); or a modification atposition 428 and/or 433 (e.g., H/L/R/S/P/Q or K) and/or 434 (e.g., A, W,H, F or Y [N434A, N434W, N434H, N434F or N434Y]); or a modification atposition 250 and/or 428; or a modification at position 307 or 308 (e.g.,308F, V308F), and 434. In one embodiment, the modification comprises a428L (e.g., M428L) and 434S (e.g., N434S) modification; a 428L, 259I(e.g., V259I), and 308F (e.g., V308F) modification; a 433K_(D) (e.g.,H433K) and a 434 (e.g., 434Y) modification; a 252, 254, and 256 (e.g.,252Y, 254T, and 256E) modification; a 250Q and 428L modification (e.g.,T250Q and M428L); and a 307 and/or 308 modification (e.g., 308F or308P). In yet another embodiment, the modification comprises a 265A(e.g., D265A) and/or a 297A (e.g., N297A) modification.

For example, the present invention includes anti-IL36R antigen-bindingproteins, e.g., antibodies or antigen-binding fragments, comprising anFc domain comprising one or more pairs or groups of mutations selectedfrom the group consisting of: 250Q and 248L (e.g., T250Q and M248L);252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g.,M428L and N434S); 257I and 311I (e.g., P257I and Q311I); 257I and 434H(e.g., P257I and N434H); 376V and 434H (e.g., D376V and N434H); 307A,380A and 434A (e.g., T307A, E380A and N434A); and 433K_(D) and 434F(e.g., H433K_(D) and N434F).

Anti-IL36R antigen-binding proteins, e.g., antibodies andantigen-binding fragments thereof, that comprise a V_(H) and/or V_(L) asset forth herein comprising any possible combinations of the foregoingFc domain mutations, are contemplated within the scope of the presentinvention.

The present invention also includes anti-IL36R antigen-binding proteins,antibodies or antigen-binding fragments, comprising a V_(H) set forthherein and a chimeric heavy chain constant (C_(H)) region, wherein thechimeric C_(H) region comprises segments derived from the C_(H) regionsof more than one immunoglobulin isotype. For example, the antibodies ofthe invention may comprise a chimeric C_(H) region comprising part orall of a C_(H)2 domain derived from a human IgG1, human IgG2 or humanIgG4 molecule, combined with part or all of a C_(H)3 domain derived froma human IgG1, human IgG2 or human IgG4 molecule. According to certainembodiments, the antibodies of the invention comprise a chimeric C_(H)region having a chimeric hinge region. For example, a chimeric hinge maycomprise an “upper hinge” amino acid sequence (amino acid residues frompositions 216 to 227 according to EU numbering) derived from a humanIgG1, a human IgG2 or a human IgG4 hinge region, combined with a “lowerhinge” sequence (amino acid residues from positions 228 to 236 accordingto EU numbering) derived from a human IgG1, a human IgG2 or a human IgG4hinge region. According to certain embodiments, the chimeric hingeregion comprises amino acid residues derived from a human IgG1 or ahuman IgG4 upper hinge and amino acid residues derived from a human IgG2lower hinge. An antibody comprising a chimeric C_(H) region as describedherein may, in certain embodiments, exhibit modified Fc effectorfunctions without adversely affecting the therapeutic or pharmacokineticproperties of the antibody. (See, e.g., WO2014/022540).

Multispecific Antigen-Binding Proteins

The present invention includes anti-IL36R antigen-binding proteins,e.g., antibodies and antigen-binding fragments thereof, as well asmethods of use thereof and methods of making such antigen-bindingproteins. The term “anti-IL36R” antigen-binding protein, e.g.,antibodies or antigen-binding fragments, includes multispecific (e.g.,bispecific or biparatopic) molecules that include at least one firstantigen-binding domain that specifically binds to IL36R (e.g., IL1RL2)(e.g., an antigen-binding domain from H4H14699P2; H4H14700P2;H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2;H4H14734P2; H4H14757P; H4H14758P or H4H14760P2) and at least one secondantigen-binding domain that binds to a different antigen or to anepitope in IL36R which is different from that of the firstantigen-binding domain (e.g., IL23-p19, IL12/IL23-p40, TNFalpha, IL-22,MADCAM, a4b7, CCR9, and/or CCR6). In an embodiment of the invention, thefirst and second epitopes overlap. In another embodiment of theinvention, the first and second epitopes do not overlap.

“H4H14699P2”; “H4H14700P2”; “H4H4706P2”; “H4H14708P2”; “H4H14709P”;“H4H14728P”; “H4H14731P”; “H4H14732P2”; “H4H14734P2”; “H4H14757P”;“H4H14758P” or “H4H14760P2” includes a multispecific molecules, e.g.,antibodies or antigen-binding fragments, that include the HCDRs andLCDRs, V_(H) and V_(L), or HC and LC of H4H14699P2; H4H14700P2;H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2;H4H14734P2; H4H14757P; H4H14758P or H4H14760P2, respectively and one ormore antigen-binding domains that bind to a different epitope.

In an embodiment of the invention, an antigen-binding domain that bindsspecifically to IL36R (e.g., IL1RL2), which may be included in amultispecific molecule, comprises:

(1)

(i) a heavy chain variable domain (V_(H)) sequence that comprisesCDR-H1, CDR-H2 and CDR-H3 from an immunoglobulin heavy chain selectedfrom: H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P andH4H14760P2, and

(ii) a light chain variable domain (V_(L)) sequence that comprisesCDR-L1, CDR-L2 and CDR-L3 from an immunoglobulin heavy chain selectedfrom: H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P andH4H14760P2, respectively;

or,(2)

(i) a heavy chain variable domain (V_(H)) sequence selected from:H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P and H4H14760P2;and

(ii) a light chain variable domain (V_(L)) sequence selected from:H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P and H4H14760P2,respectively; and

one or more antigen-binding domains that bind to a different epitope.

In an embodiment of the invention, the multispecific antibody orfragment includes more than two different binding specificities (e.g., atrispecific molecule), for example, one or more additionalantigen-binding domains which are the same or different from the firstand/or second antigen-binding domain.

In one embodiment of the invention, a bispecific antigen-bindingfragment comprises a first scFv (e.g., comprising V_(H) and V_(L) ofH4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P and H4H14760P2)having binding specificity for a first epitope (e.g., IL36R) and asecond scFv having binding specificity for a second, different epitope.For example, in an embodiment of the invention, the first and secondscFv are tethered with a linker, e.g., a peptide linker (e.g., a GSlinker such as (GGGGS)_(n) (SEQ ID NO: 177) wherein n is, for example,1, 2, 3, 4, 5, 6, 7, 8, 9 or 10).

Other bispecific antigen-binding fragments include an F(ab)₂ of abispecific IgG antibody which comprises the heavy and light chain CDRsof H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P and H4H14760P2and of another antibody that binds to a different epitope.

Immunoconjugates

The invention encompasses anti-IL36R antigen-binding proteins, e.g.,antibodies or antigen-binding fragments, conjugated to another moiety,e.g., a therapeutic moiety (an “immunoconjugate”). In an embodiment ofthe invention, an anti-IL36R antigen-binding protein, e.g., antibody orantigen-binding fragment, is conjugated to any of the furthertherapeutic agents set forth herein. As used herein, the term“immunoconjugate” refers to an antigen-binding protein, e.g., anantibody or antigen-binding fragment, which is chemically orbiologically linked to another antigen-binding protein, a radioactiveagent, a reporter moiety, an enzyme, a peptide, a protein or atherapeutic agent.

Therapeutic and Prophylactic Methods

The present invention provides methods for treating or preventing anIL-36R-mediated disease by administering a therapeutically effectiveamount of anti-IL36R antigen-binding protein, e.g., antibody orantigen-binding fragment, (e.g., H4H14699P2; H4H14700P2; H4H14706P2;H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2;H4H14757P; H4H14758P or H4H14760P2) to a subject (e.g., a human) in needof such treatment or prevention.

“Treat” or “treating” means to administer an anti-IL36R antigen-bindingprotein, e.g., antibody or antigen-binding fragment of the presentinvention (e.g., H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P;H4H14758P or H4H14760P2), to a subject, having an IL36R-mediateddisease, such that one or more signs and/or symptoms and/or clinicalindicia of the IL36R-mediated disease regresses or is eliminated and/orthe progression thereof is inhibited (e.g., the disease in the subjectis stabilized, reduced or eliminated).

“Preventing” an IL36R-mediated disease means to administer anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragment ofthe present invention (e.g., H4H14699P2; H4H14700P2; H4H14706P2;H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2;H4H14757P; H4H14758P or H4H14760P2), to a subject, prior tomanifestation of the disease in the body of the subject.

Interleukin IL-36RN is an IL-1 cytokine family member that antagonizesthe proinflammatory signals of IL-36alpha, IL-36beta and IL-36gamma atthe IL-36R.

An IL-36R-mediated disease is any disease which is caused or exacerbatedby an activity of IL-36R (e.g., activation of downstream inflammatorysignaling via NFκB and MAP kinases due to receptor binding of ligandsuch as IL36γ, IL36γ and/or IL36γ), for example, due to a deficiency inan IL36R antagonist (e.g., IL-36RN). In an embodiment of the invention,a mutation in IL36RN underlies the IL-36R-mediated disease. An exampleof such a disease is an autoimmune and/or inflammatory disorder. Inembodiment of the invention, the IL-36R-mediated disease treated with ananti-IL36R antigen-binding protein is an inflammatory disorder, anautoimmune disorder, deficiency of interleukin IL-36 receptor antagonist(DITRA) syndrome, impetigo herpetiformis, acrodermatitis, a skinneutrophilic pustular disease, psoriasis, a pustular disease,generalized pustular psoriasis (GPP; e.g., familial or sporadic),psoriasis vulgaris/plaque psoriasis, palmoplantar pustular psoriasis(PPPP), palmoplantar pustulosis (PPP), colitis, inflammatory boweldisease, ulcerative colitis, Crohn's disease, chemically-inducedcolitis, inflammation, airway inflammation (e.g., neutrophilic airwayinflammation, COPD (chronic obstructive pulmonary disease) or asthma),joint inflammation (e.g., ankylosing spondylitis, rheumatoid arthritisor psoriatic arthritis), kidney inflammation, alopecia areata, skininflammation (e.g., chemically-induced skin inflammation, psoriasis,pustular psoriasis, generalized pustular psoriasis, palmoplantarpustulosis, palmo-plantar pustular psoriasis, psoriasis vulgaris orpsoriatic skin lesions), acanthosis, hyperkeratosis, kindler syndrome,systemic lupus erythematosus (SLE), nephrotic syndrome, ANCA(anti-neutrophil cytoplasmic antibody)-associated vasculopathies,tubulointerstitial lesions and glomerulonephritis.

An inflammatory disorder is a disorder characterized by uncontrolled orunwanted inflammation which may cause destruction of healthy tissue.

An autoimmune disorder is a condition in which one's immune systemmistakenly attacks one's own body.

Impetigo herpetiformis (IH) is among rare dermatosis of pregnancy, whichis currently considered as a form of generalized pustular psoriasis. Inan embodiment of the invention, a mutation in IL36RN underlies the IH.

Acrodermatitis is a skin condition that may affect children, e.g.,between the ages of 3 months and 15 years, which is characterized byitchy red or purple blisters on the body, bloated abdomen, fever, andswollen, sore lymph nodes. The cause of acrodermatitis may be viral.Mutations of IL-36 receptor antagonists (e.g., IL-36Ra) are present in ahigh proportion of patients with GPP and acrodermatitis continua. In anembodiment of the invention, a mutation in IL36RN underlies theacrodermatitis.

Psoriasis is an autoimmune disease that causes skin plaques, which areitchy or sore patches of thick, red, dry skin. The most common form ofpsoriasis is psoriasis vulgaris (plaque psoriasis) which ischaracterized by well-defined plaques of red raised skin that can appearon any area of skin, including the knees, elbows, scalp and trunk. Aflaky silvery white buildup on top of the plaques is called scale; it iscomposed of dead skin cells. This scale comes loose and sheds constantlyfrom the plaques. Skin symptoms include pain, itching and cracking.

Generalized pustular psoriasis (GPP) is a severe form of psoriasis.Individuals with GPP typically have repeated episodes in which largeareas of skin become red and inflamed and develop small pus-filledblisters (pustules). A portion of subjects with GPP suffer from plaques.The skin problems can be accompanied by fever, extreme tiredness(fatigue), muscle weakness, an increased number of white blood cells,and other signs of inflammation throughout the body (systemicinflammation). IL-36 cytokine appears to play a role in the developmentof GPP. In an embodiment of the invention, a mutation in IL36RNunderlies the GPP.

Palmoplantar pustular psoriasis (PPPP; 4P) is a form of localizedpustular psoriasis characterized by plaques and pustules occurring onpalmar and plantar surfaces of the skin. PPPP may be associated withhomozygous or compound heterozygous IL36RN gene mutations leading toaberrations in IL-36R antagonist function. In an embodiment of theinvention, a mutation in IL36RN underlies the PPPP.

Palmoplantar pustulosis (PPP; 3P) is an immune-mediated disorder thatcauses blister-like pustules to show up on the palms of your hands andthe soles of your feet. Generally, subjects with PPP do not suffer fromplaques. In an embodiment of the invention, a mutation in IL36RNunderlies the PPP.

Deficiency of interleukin IL-36 receptor antagonist (DITRA) syndrome isa rare autosomal recessive disease caused by mutations in IL36RN. DITRAis a rare, genetic, auto-inflammatory syndrome with immune deficiencydisease characterized by recurrent and severe flares of generalizedpustular psoriasis associated with high fever, asthenia, and systemicinflammation, due to IL36R antagonist deficiency. Psoriatic nail changes(e.g., pitting and onychomadesis) and ichthyosis may occasionally beassociated. See Marrakchi et al., New Engl J. Med. 365(7): 620-628(2011). In an embodiment of the invention, a mutation in IL36RNunderlies the DITRA.

An inflammatory disease is a condition characterized by abnormalinflammation at one or more sites within the body of a subject. Anautoimmune disease is a condition characterized by the abnormal attackof the subject's body tissue by the subject's own immune system.

ANCA-associated vasculopathies (AAV) are inflammatory disorders thatinclude Granulomatosis with polyangiitis (formerly Wegener's),microscopic polyangitis, and EGPA/Churg Strauss. These conditions arecharacterized by chronic inflammation leading to blockages of bloodvessels and diminished blood flow to vital organs like the kidney.

Inflammatory bowel disease (IBD) is a term that includes two conditions(Crohn's disease and ulcerative colitis) that are characterized bychronic inflammation of the gastrointestinal (GI) tract.

Neutrophilic airway inflammation is inflammation of the airway which ismediated by the influx of neutrophils into the lungs. Signs and symptomsof neutrophilic airway inflammation include asthma and wheezing.

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatorylung disease that causes obstructed airflow from the lungs. Signs andsymptoms include breathing difficulty, cough, mucus (sputum) productionand wheezing.

Ankylosing spondylitis (AS) is a disease characterized by long terminflammation of the spine (e.g., the sacroiliac (SI) joints and theaxial skeleton). Over time, AS can cause some of the vertebrae in yourspine to fuse. Symptoms include pain and stiffness in your lower backand hips.

Rheumatoid arthritis is an autoimmune condition characterized by jointinflammation. Symptoms include tender, warm, swollen joints; jointstiffness, fatigue, fever and weight loss.

Psoriatic arthritis is a form of arthritis that affects some people whohave psoriasis. Symptoms can include swollen fingers and toes, foot painand lower back pain.

Alopecia areata is spot baldness characterized by small bald patches onthe body.

Acanthosis is diffuse epidermal thickening (hyperplasia) of the stratumspinosum (prickle cell layer) of the skin which may appear to be darkerthan other skin. Hyperkeratosis is a thickening of the outer layer ofthe skin.

Hyperkeratosis is the thickening of skin often due to irritation fromthe sun, chemicals or frequent friction or pressure. The skin thickeningtypically occurs in the outer layer of the skin, which contains a tough,protective protein called keratin.

Kindler syndrome is an autosomal recessive genodermatosis characterizedby congenital acral skin blistering, photosensitivity, progressivepoikiloderma, and diffuse cutaneous atrophy. Mucosal manifestations arecommon, with frequent involvement of the oral mucosa, gingiva, andgastrointestinal tract.

Systemic lupus erythematosus (SLE) is an autoimmune disease. In thisdisease, the body's immune system mistakenly attacks healthy tissue. SLEcan affect the skin, joints, kidneys, brain, and other organs.

Nephrotic syndrome is a kidney disorder that causes your body to excretetoo much protein in your urine. Nephrotic syndrome is typically causedby damage to the clusters of small blood vessels in the kidneys thatfilter waste and excess water from your blood. Nephrotic syndromesymptoms may include swelling (edema), particularly in the feet andankles, foamy urine, weight gain (from fluid retention), fatigue andloss of appetite.

Glomerulonephritis is inflammation of kidney glomeruli. Symptoms includeink or cola-colored urine from red blood cells in your urine(hematuria), foamy urine (due to proteinuria), high blood pressure(hypertension), fluid retention (edema).

An effective or therapeutically effective dose of anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragment(e.g., H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P;H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P orH4H14760P2), for treating or preventing an IL-36R-mediated diseaserefers to the amount of the antibody or fragment sufficient to alleviateone or more of the clinical indicia, signs and/or symptoms of thedisease in the treated subject, whether by inducing the regression orelimination of such indicia, signs and/or symptoms or by inhibiting theprogression of such indicia, signs and/or symptoms. The dose amount mayvary depending upon the age and the size of a subject to beadministered, target disease, conditions, route of administration, andthe like. In an embodiment of the invention, an effective ortherapeutically effective dose of antibody or antigen-binding fragmentthereof of the present invention, for treating or preventing IL36Rmediated disease, e.g., in an adult human subject, is about 1 mg/kg ormore, e.g., about 1 mg/kg to about 25 mg/kg. Depending on the severityof the infection, the frequency and the duration of the treatment can beadjusted. In certain embodiments, the antigen-binding protein of thepresent invention can be administered at an initial dose, followed byone or more secondary doses. In certain embodiments, the initial dosemay be followed by administration of a second or a plurality ofsubsequent doses of antigen-binding protein in an amount that can beapproximately the same or less than that of the initial dose, whereinthe subsequent doses are separated by at least 1 day to 3 days; at leastone week, at least 2 weeks; at least 3 weeks; at least 4 weeks; at least5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks.

As used herein, the term “subject” refers to a mammal (e.g., rat, mouse,cat, dog, cow, sheep, horse, goat, rabbit), preferably a human, forexample, in need of prevention and/or treatment of an IL-36R-mediateddisease. The subject may have an IL-36R-mediated disease or bepredisposed to developing such a disease. In an embodiment of theinvention, the subject has a homozygous or heterozygous IL36RN mutationgenotype.

The present invention encompasses methods for administering ananti-IL36R antigen-binding protein to a subject at risk of developing anIL36R-mediated disease. For example, in an embodiment of the invention,the disease is a skin inflammatory disease or colon inflammatorydisease. Example 5 herein demonstrated that skin inflammation diseasescould be prevented in a DITRA-like mouse model prior to exposure toimiquimod and the development of skin inflammation symptoms. In anembodiment of the invention, an IL36R-mediated disease (e.g., skininflammation) is prevented by administration of a prophylactic dose ofantigen-binding protein to a subject prior to any clinically significantinflammation, e.g., skin inflammation or any increase ininflammation-induced skin thickness, in total pathology score (asdiscussed herein) or in the presence of pro-inflammatory cytokines, suchas KC-GRO, IL-6, IL-1beta or TNFalpha, in the skin. In an embodiment ofthe invention, a dose of anti-IL36R antigen-binding protein of theinvention for preventing an IL36R-mediated disease is from about 1 mg/kgto about 10 mg/kg.

Combinations and Pharmaceutical Compositions

The present invention provides compositions that include anti-IL36Rantigen-binding proteins and one or more ingredients; as well as methodsof use thereof and methods of making such compositions.

To prepare pharmaceutical compositions of the anti-IL36R antigen-bindingproteins, e.g., antibodies and antigen-binding fragments thereof (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2),antigen-binding protein is admixed with a pharmaceutically acceptablecarrier or excipient. See, e.g., Remington's Pharmaceutical Sciences andU.S. Pharmacopeia: National Formulary, Mack Publishing Company, Easton,Pa. (1984); Hardman, et al. (2001) Goodman and Gilman's ThePharmacological Basis of Therapeutics, McGraw-Hill, New York, N.Y.;Gennaro (2000) Remington: The Science and Practice of Pharmacy,Lippincott, Williams, and Wilkins, New York, N.Y.; Avis, et al. (eds.)(1993) Pharmaceutical Dosage Forms: Parenteral Medications, MarcelDekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms:Tablets, Marcel Dekker, NY: Lieberman, et al. (eds.) (1990)Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weinerand Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc.,New York, N.Y. In an embodiment of the invention, the pharmaceuticalcomposition is sterile. Such compositions are part of the presentinvention.

Pharmaceutical compositions of the present invention includepharmaceutically acceptable carriers, diluents, excipients and/orstabilizers, such as, for example, water, buffering agents, stabilizingagents, preservatives, isotonifiers, non-ionic detergents, antioxidantsand/or other miscellaneous additives.

The scope of the present invention includes desiccated, e.g.,freeze-dried, compositions comprising an anti-IL36R antigen-bindingprotein, e.g., antibody or antigen-binding fragment thereof (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2),or a pharmaceutical composition thereof that includes a pharmaceuticallyacceptable carrier but substantially lacks water.

In a further embodiment of the invention, a further therapeutic agentthat is administered to a subject in association with an anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragmentthereof (e.g., H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P;H4H14758P or H4H14760P2), disclosed herein is administered to thesubject in accordance with the Physicians' Desk Reference 2003 (ThomsonHealthcare; 57^(th) edition (Nov. 1, 2002)).

The mode of administration of an antigen-binding protein or compositionthereof can vary. Routes of administration include oral, rectal,transmucosal, intestinal, parenteral; intramuscular, subcutaneous,intradermal, intramedullary, intrathecal, direct intraventricular,intravenous, intraperitoneal, intranasal, intraocular, inhalation,insufflation, topical, cutaneous, transdermal or intra-arterial.

The present invention provides methods for administering an anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragmentthereof (e.g., H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P;H4H14758P or H4H14760P2) to a subject, comprising introducing theprotein or a pharmaceutical composition or combination thereof into thebody of the subject. For example, in an embodiment of the invention, themethod comprises piercing the body of the subject, e.g., with a needleof a syringe, and injecting the antigen-binding protein or apharmaceutical composition or combination thereof into the body of thesubject, e.g., into the vein, artery, tumor, muscular tissue or subcutisof the subject.

The present invention provides a vessel (e.g., a plastic or glass vial,e.g., with a cap or a chromatography column, hollow bore needle or asyringe cylinder) comprising any of the anti-IL36R antigen-bindingproteins, e.g., antibodies or antigen-binding fragments thereof (e.g.,H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2; H4H14709P; H4H14728P;H4H14731P; H4H14732P2; H4H14734P2; H4H14757P; H4H14758P or H4H14760P2),or a pharmaceutical composition comprising a pharmaceutically acceptablecarrier or combination thereof.

The present invention includes combinations including an anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragmentthereof of the present invention (e.g., H4H14699P2; H4H14700P2;H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2;H4H14734P2; H4H14757P; H4H14758P or H4H14760P2), in association with oneor more further therapeutic agents. The anti-IL36R antigen-bindingprotein and the further therapeutic agent can be in a single compositionor in separate compositions. For example, in an embodiment of theinvention, the further therapeutic agent is an anti-inflammatory drug.In an embodiment of the invention, the further therapeutic agent isanother anti-IL35R antigen-binding protein, an IL17 inhibitor, anIL23p19 inhibitor, an IL12p40 inhibitor, guselkumab, ustekinumab,brodalumab, ixekizumab, secukinumab, an anti-TNFalpha antibody orantigen-binding fragment thereof, one or more human TNF receptors orfragments thereof linked to an immunoglobulin such as an Fc portion of ahuman IgG1, infliximab, adalimumab, etanercept, dupilumab, sarilumab,tocilizumab, golimumab, abatacept, tofacitinib, abatacept, anon-steroidal anti-inflammatory drug (NSAID), ibuprofen, naproxen,acetaminophen, aspirin, celecoxib, cyclophosphamide, methotrexate, acorticosteroid, cortisone or prednisone.

Methods for treating or preventing an IL-36-mediated disease in asubject in need of said treatment or prevention by administering ananti-IL36R antigen-binding protein, e.g., H4H14699P2; H4H14700P2;H4H14706P2; H4H14708P2; H4H14709P; H4H14728P; H4H14731P; H4H14732P2;H4H14734P2; H4H14757P; H4H14758P or H4H14760P2, in association with afurther therapeutic agent are part of the present invention.

The term “in association with” indicates that components, an anti-IL36Rantigen-binding protein, e.g., antibody or antigen-binding fragmentthereof of the present invention, along with another agent such asmethotrexate, can be formulated into a single composition, e.g., forsimultaneous delivery, or formulated separately into two or morecompositions (e.g., a kit including each component). Each component canbe administered to a subject at a different time than when the othercomponent is administered; for example, each administration may be givennon-simultaneously (e.g., separately or sequentially) at intervals overa given period of time. Moreover, the separate components may beadministered to a subject by the same or by a different route.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the methods and compositions of the invention, and are notintended to limit the scope of what the inventors regard as theirinvention.

Example 1: Generation of Human Antibodies that Specifically Bind toIL-36R

Anti-IL36R antibodies were obtained by immunizing a VELOCIMMUNE mouse(i.e., an engineered mouse comprising DNA encoding human immunoglobulinheavy and kappa light chain variable regions) with a DNA immunogencomprising the full length IL36R (IL-1RL2) sequence. The antibody immuneresponse was monitored by an IL36R-specific immunoassay and fully humananti-IL36R antibodies were isolated and purified. Two exemplarycomparisons between the V_(H) and V_(L) of antibodies generated as setforth herein and their respective germlines are set forth in FIG. 1 andFIG. 2.

TABLE 1 Immunoglobulin chain sequences of the present invention*Antibody VH CDR1 CDR2 CDR3 # Name DNA PEP DNA PEP DNA PEP DNA PEP 1H4H14699P2 1 2 3 4 5 6 7 8 2 H4H14700P2 17 18 19 20 21 22 23 24 3H4H14706P2 33 34 35 36 37 38 39 40 4 H4H14708P2 49 50 51 52 53 54 55 565 H4H14709P 65 66 67 68 69 70 71 72 6 H4H14728P 81 82 83 84 85 86 87 887 H4H14731P 97 98 99 100 101 102 103 104 8 H4H14732P2 113 114 115 116117 118 119 120 9 H4H14734P2 129 130 131 132 133 134 135 136 10H4H14757P 137 138 139 140 141 142 143 144 11 H4H14758P 153 154 155 156157 158 159 160 12 H4H14760P2 169 170 171 172 173 174 175 176 AntibodyVK CDR1 CDR2 CDR3 # Name DNA PEP DNA PEP DNA PEP DNA PEP 1 H4H14699P2 910 11 12 13 14 15 16 2 H4H14700P2 25 26 27 28 29 30 31 32 3 H4H14706P241 42 43 44 45 46 47 48 4 H4H14708P2 57 58 59 60 61 62 63 64 5 H4H14709P73 74 75 76 77 78 79 80 6 H4H14728P 89 90 91 92 93 94 95 96 7 H4H14731P105 106 107 108 109 110 111 112 8 H4H14732P2 121 122 123 124 125 126 127128 9 H4H14734P2 121 122 123 124 125 126 127 128 10 H4H14757P 145 146147 148 149 150 151 152 11 H4H14758P 161 162 163 164 165 166 167 168 12H4H14760P2 121 122 123 124 125 126 127 128 *Numbers corresponding toV_(H), CDR-H1, CDR-H2, CDR-H3, V_(L), CDR-L1, CDR-L2 and CDR-L3 refer toSEQ ID NOs set forth herein. “PEP” refers to an amino acid sequence;“DNA” refers to a nucleotide sequence.

SEQ ID NO: 1GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCGGCCTCTGGATTCACCTTTGATGATTATGCCATACACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATCAGTTGGAATAGTGATATCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCGTCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAATAGTCTGAGAACTGAGGACACGGCCTTGTATTACTGTGCAAAAGGATATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 2EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAIHWVRQAPGKGLEWVSVISWNSDIIGYADSVKGRFTVSRDNAKNSLYLQMNSLRTEDTALYYGAKGYNWNFFDYWGQGTLVTVSS; SEQ ID NO: 3GGA TTC ACC TTT GAT GAT TAT GCC; SEQ ID NO: 4 G F T F D D Y A;SEQ ID NO: 5 ATC AGT TGG AAT AGT GAT ATC ATA; SEQ ID NO: 6I S W N S D I I; SEQ ID NO: 7GCA AAA GGA TAT AAC TGG AAC TTC TTT GAC TAT; SEQ ID NO: 8A K G Y N W N F F D Y; SEQ ID NO: 9GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTATCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGCAGCAAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA; SEQ ID NO: 10EIVLTQSPATLSLSPGEFATLSCPASQSVSSYLAWYQQKPGQAPRLLIYNAANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIK; SEQ ID NO: 11 CAG AGT GTT AGC AGC TAC;SEQ ID NO: 12 Q S V S S Y; SEQ ID NO: 13 AAT GCA GCA; SEQ ID NO: 14N A A; SEQ ID NO: 15 CAG CAG CGT AGC AAC TGG CCT CTC ACT; SEQ ID NO: 16Q Q R S N W P L T; SEQ ID NO: 17GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAACTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATTAGTTGGAATAGTGATGTCATAGCCTATTCGGACTCTGTGAAGGGCCGCTTCACCATTTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGGGAACTGAGGACACGGCCTTATATTACTGTGCAAAAGGCCATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 18EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQTPGKGLEWVSVISWNSDVIAYSDSVKGRFTISRDNAKNSLYLQMNSLGTEDTALYYCAKGHNWNFFDYWGQGTLVTVSS; SEQ ID NO: 19GGA TTC ACC TTT GAT GAT TAT GCC; SEQ ID NO: 20 G F T F D D Y A;SEQ ID NO: 21 ATT AGT TGG AAT AGT GAT GTC ATA; SEQ ID NO: 22I S W N S D V I; SEQ ID NO: 23GCA AAA GGC CAT AAC TGG AAC TTC TTT GAC TAT; SEQ ID NO: 24A K G H N W N F F D Y; SEQ ID NO: 25GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGAGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGTAGCCAACAGGGCCACAGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCGGCCTAGAGCCTGAAGATTTTGCAGTTTATTTCTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA; SEQ ID NO: 26EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYNVANRATDIPARFSGSGSGTDFTLTISGLEPEDFAVYFCQQRSNWPLTFGGGTKVEIK; SEQ ID NO: 27 CAG AGT GTT AGC AGC TAG;SEQ ID NO: 28 Q S V S S Y; SEQ ID NO: 29 AAT GTA GCC; SEQ ID NO: 30N V A; SEQ ID NO: 31 CAG CAG CGT AGC AAC TGG CCT CTC ACT; SEQ ID NO: 32Q Q R S N W P L T; SEQ ID NO: 33GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTTGATGATTATGCCATACACTGGGTCCGGCAATCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATCAGTTGGAATAGTGATGTCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAGATGAATAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAAGGATATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 34EVQLVESGGGLVQPGRSLRLSCTASGFTFDDYAIHWVRQSPGKGLEWVSVISWNSDVIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKGYNWNFFDYWGQGTLVTVSS; SEQ ID NO: 35GGA TTC ACC TTT GAT GAT TAT GCC; SEQ ID NO: 36 G F T F D D Y A;SEQ ID NO: 37 ATC AGT TGG AAT AGT GAT GTC ATA; SEQ ID NO: 38I S W N S D V I; SEQ ID NO: 39GCA AAA GGA TAT AAC TGG AAC TTC TTT GAC TAT; SEQ ID NO: 40A K G Y N W N F F D Y; SEQ ID NO: 41GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTATCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGCAGCAAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA; SEQ ID NO: 42EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYNAANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIK; SEQ ID NO: 43 CAG AGT GTT AGC AGC TAG;SEQ ID NO: 44 Q S V S S Y; SEQ ID NO: 45 AAT GCA GCA; SEQ ID NO: 46N A A; SEQ ID NO: 47 CAG CAG CGT AGC AAC TGG CCT CTC ACT; SEQ ID NO: 48Q Q R S N W P L T; SEQ ID NO: 49GAAGTGCAGCTGGTGGAGTCTGGGGGAGACTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAATGGGTCTCAGTTATTAGTTGGAATAGTGATGTCATAGCCTATTCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGAACTGAGGACACGGCCTTATATTACTGTACAAAAGGCCATAAGTGGAGCTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 50EVQLVESGGDLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSVISWNSDVIAYSDSVKGRFTISRDNAKNSLYLQMNSLRTEDTALYYCTKGHKWSFFDYWGQGTLVTVSS; SEQ ID NO: 51GGA TTC ACC TTT GAT GAT TAT GCC; SEQ ID NO: 52 G F T F D D Y A;SEQ ID NO: 53 ATT AGT TGG AAT AGT GAT GTC ATA; SEQ ID NO: 54I S W N S D V I; SEQ ID NO: 55ACA AAA GGC CAT AAG TGG AGC TTC TTT GAC TAT; SEQ ID NO: 56T K G H K W S F F D Y; SEQ ID NO: 57GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTATTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGACTCCTCATCTTTAATGTAGCCAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA; SEQ ID NO: 58EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIFNVANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIK; SEQ ID NO: 59 CAG AGT ATT AGC AGC TAC;SEQ ID NO: 60 Q S I S S Y; SEQ ID NO: 61 AAT GTA GCC; SEQ ID NO: 62N V A; SEQ ID NO: 63 CAG CAG CGT AGC AAC TGG CCT CTC ACT; SEQ ID NO: 64Q Q R S N W P L T; SEQ ID NO: 65GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTTCAGCCTGGGGGGTCCCTGAGACTCTCCTGCGCAGCCTCTGGATTCACCTTTAGCGACTATGCCATGAGCTGGGTCCGCCAGGCTCCGGGGAAGGGGCTGGAGTGGGTCTCAGGTATTAGTGGAAATGGTGGTGACACATACTACGGAGACTTCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGGCGAGGACACGGCCGCATATTTCTGTGTGATAGATCTTGACTATTGGGGTCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 66EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYAMSWVRQAPGKGLEWVSGISGNGGDTYYGDFVKGRFTISRDNSKNTLYLQMNSLRGEDTAAYFCVIDLDYWGQGTLVTVSS; SEQ ID NO: 67GGA TTC ACC TTT AGC GAC TAT GCC; SEQ ID NO: 68 G F T F S D Y A;SEQ ID NO: 69 ATT AGT GGA AAT GGT GGT GAC ACA; SEQ ID NO: 70I S G N G G D T; SEQ ID NO: 71 GTG ATA GAT CTT GAC TAT; SEQ ID NO: 72V I D L D Y; SEQ ID NO: 73GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGAAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAACAGAAACCAGGAAAAGCCCCTAGGCTCCTGATCTATAAGGCGTCTATTTTAGGAGATGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCTACTTATTACTGCCACCAGTATAATAGTTATTTGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA; SEQ ID NO: 74DIQMTQSPSTLSASEGDRVTITCRASQSISSWLAWYQQKPGKAPRLLIYKASILGDGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCHQYNSYLWTFGQGTKVEIK; SEQ ID NO: 75 CAG AGT ATT AGT AGC TGG;SEQ ID NO: 76 Q S I S S W; SEQ ID NO: 77 AAG GCG TCT; SEQ ID NO: 78K A S; SEQ ID NO: 79 CAC CAG TAT AAT AGT TAT TTG TGG ACG; SEQ ID NO: 80H Q Y N S Y L W T; SEQ ID NO: 81CAGGTGCAGCTGGAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGCTGATTACTATTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGATCCATCTATTATACTGGGAGTACTTACTACAACCCGTCCCTCAAGAGTCGACTTACCATATCAATAGACACGTCTGAGAACCAGTTCTCTTTGAAACTGACCTCTCTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGCGAGGAGGCTAACTGGGGATCCCACTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 82QVQLQESGPGLVKPSQTLSLTCTVSGGSISSADYYWSWIRQHPGKGLEWIGSIYYTGSTYYNPSLKSRLTISIDTSENQFSLKLTSLTAADTAVYYCASEEANWGSHFDSWGQGTLVTVSS; SEQ ID NO: 83GGT GGC TCC ATC AGC AGT GCT GAT TAC TAT; SEQ ID NO: 84G G S I S S A D Y Y; SEQ ID NO: 85 ATC TAT TAT ACT GGG AGT ACT;SEQ ID NO: 86 I Y Y T G S T; SEQ ID NO: 87GCG AGC GAG GAG GCT AAC TGG GGA TCC CAC TTT GAC TCC; SEQ ID NO: 88A S E E A N W G S H F D S; SEQ ID NO: 89GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGACAACTTTTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCATCTTACTACTGTCAACATAGTCACAGTGCCCATCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAA; SEQ ID NO: 90DIQMTQSPSSLSASVGDRVTITCRASQSIDNFLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFASYYCQHSKSAHPITFGQGTRLEIK; SEQ ID NO: 91 CAG AGC ATT GAC AAC TTT;SEQ ID NO: 92 Q S I D N F; SEQ ID NO: 93 GCT GGA TCC; SEQ ID NO: 94A A S; SEQ ID NO: 95 CAA CAT AGT CAC AGT GCC CAT CCG ATC ACC;SEQ ID NO: 96 Q H S H S A H P I T; SEQ ID NO: 97CAGCTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTAGTAATTACTACTGGGGCTGGATCCGCCAGCCCCCAGGGAAGAGACTGGAGTGGATTGGGAGTATCTATTATAGTGGGAGCACCTACTACAACCCGTCCCTCAAGACTCGAGTCACCATATCCGTAGACACGTCCAAGAATCAGTTCTCCCTGAAGCTGACCTCTGTGACCGCCGCAGACACGGCTGTGTATTACTGTGCGAGAGAGGAAGCAGCAGCTTTGACGCACTTTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 98QLQLQESGPGLVKPSETLSLTCTVSGGSISSSNYYWGWIRQPPGKRLEWIGSIYYSGSTYYNPSLKTRVTISVDTSKNQFSLKLTSVTAADTAVYYCAREEAAALTHFDFWGQGTLVTVSS; SEQ ID NO: 99GGT GGC TCC ATC AGC AGT AGT AAT TAG TAG; SEQ ID NO: 100G G S I S S S N Y Y; SEQ ID NO: 101 ATC TAT TAT AGT GGG AGC ACC;SEQ ID NO: 102 I Y Y S G S T; SEQ ID NO: 103GCG AGA GAG GAA GCA GCA GCT TTG ACG CAC TTT GAC TTC; SEQ ID NO: 104A R E E A A A L T H F D F; SEQ ID NO: 105GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAACTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTGCTGCATCCAGTTTACAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACATAGTCACAGTTCCCATCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAA; SEQ ID NO: 106DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIFAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHSKSSHPITFGQGTRLEIK; SEQ ID NO: 107 CAG AGC ATT AGC AAC TAT;SEQ ID NO: 108 Q S I S N Y; SEQ ID NO: 109 GCT GCA TCC; SEQ ID NO: 110A A S ; SEQ ID NO: 111 CAA CAT AGT CAC AGT TCC CAT CCG ATC ACC;SEQ ID NO: 112 Q H S H S S H P I T; SEQ ID NO: 113GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAATTGGGCTGGTTATAACATAGACTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAAGATATGCGTGGATTCAGTTATGGTTTCCCCTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA; SEQ ID NO: 114EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSGINWAGYNIDYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDMRGFSYGFPFDYWGQGTLVTVSS; SEQ ID NO: 115GGA TTC ACC TTT GAT GAT TAT GCC, SEQ ID NO: 116 G F T F D D Y A;SEQ ID NO: 117 ATT AAT TGG GCT GGT TAT AAC ATA; SEQ ID NO: 118I N W A G Y N I; SEQ ID NO: 119GCA AAA GAT ATG CGT GGA TTC AGT TAT GGT TTC CCC TTT GAC TAC;SEQ ID NO: 120 A K D M R G F S Y G F P F D Y; SEQ ID NO: 121GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCGTCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAA; SEQ ID NO: 122DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK; SEQ ID NO: 123 CAG AGC ATT AGC AGC TAT;SEQ ID NO: 124 Q S I S S Y; SEQ ID NO: 125 GCT GCA TCC; SEQ ID NO: 126A A S; SEQ ID NO: 127 CAA CAG AGT TAG AGT ACC CCT CCG ATC ACC;SEQ ID NO: 128 Q Q S Y S T P P I T; SEQ ID NO: 129GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTAAAGCCGGGGGGGTCCCTTAGACTCTCCTGTGCAGCCTCTGGATTTATTTTCAGTAACGCCTGGATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGACTGGCGTGGGTTGGCCGTATTAAAACCGAAACTGATGGTGGGACAACAGACTACGCTGCACCCGTAAAAGGCAGATTCACCATCTCAAGAGATGACTCAAAAAACACGCTGTATCTGCAAATGAACAGCGTGAAAACCGAGGACACAGCCGTGTATTACTGTACAGGGGGATACAGCTATGGTGACGATAGCAGCAGCTGGAACGAGGGCTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA;SEQ ID NO: 130EVQLVESGGGLVKPGGSLRLSCAASGFIFSNAWMNWVRQAPGKGLAWVGRIKTETDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSVKTEDTAVYYCTGGYSYGDDSSSWNEGYYYYGMDVWGQGTTVTVSS; SEQ ID NO: 131GGA TTT ATT TTC AGT AAC GCC TGG; SEQ ID NO: 132 G F I F S N A W;SEQ ID NO: 133 ATT AAA ACC GAA ACT GAT GGT GGG ACA ACA; SEQ ID NO: 134I K T E T D G G T T; SEQ ID NO: 135ACA GGG GGA TAG AGC TAT GGT GAC GAT AGC AGC AGC TGG AAC GAG GGC TAC TAG TAC TAGGGT ATG GAC GTC; SEQ ID NO: 136T G G Y S Y G D D S S S W N E G Y Y Y Y G M D V; SEQ ID NO: 137GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTCGTTGGAATGGTGGTAGTATAGGCTATGTGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAAGTCCCTGCATCTGCAAATGAACAGTCTAAAAACTGAGGACACGGCCTTGTATTACTGTGCAAAAGATATAGGCGATATTTTGACTGGTTTTTATGGAGAATACGGAATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA;SEQ ID NO: 138EVQLVESGGGLVQPGRSLRLSCTASGFTFDDYAMHWVRQAPGKGLEWVSGIRWNGGSIGYVDSVKGRFTISRDNAKKSLHLQMNSLKTEDTALYYCAKDIGDILTGFYGEYGMDVWGQGTTVTVSS; SEQ ID NO: 139GGA TTC ACC TTT GAT GAT TAT GCC; SEQ ID NO: 140 G F T F D D Y A;SEQ ID NO: 141 ATT CGT TGG AAT GGT GGT AGT ATA; SEQ ID NO: 142I R W N G G S I; SEQ ID NO: 143GCA AAA GAT ATA GGC GAT ATT TTG ACT GGT TTT TAT GGA GAA TAG GGA ATG GAC GTC;SEQ ID NO: 144 A K D I G D I L T G F Y G E Y G M D V; SEQ ID NO: 145GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGAAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAAGCAGGGAAAGCCCCTAACCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACATTATCCCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA; SEQ ID NO: 146DIQMTQSPSSLSASEGDRVTITCRASQSISSYLNWYQQKAGKAPNLLIYAASSLQSGVPSRFSGSGSGTEYTLTISSLQPEDFATYYCQQSYIIPYTFGQGTKLEIK; SEQ ID NO: 147 CAG AGC ATT AGC AGC TAT;SEQ ID NO: 148 Q S I S S Y; SEQ ID NO: 149 GCT GCA TCC; SEQ ID NO: 150A A S; SEQ ID NO: 151 CAA CAG AGT TAG ATT ATC CCG TAG ACT;SEQ ID NO: 152 Q Q S Y I I P Y T; SEQ ID NO: 153GAAGTGCAGCTGGTGGAGTCTGGGGGAGGGTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAAGTGTTAGGTGGAATGGTGGTATTATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGACCTGAGGACACGGCCCTCTATTACTGTGCAAAAGATATAGGCGATGTTTTGACTGGTTATTATGGAGAATACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA;SEQ ID NO: 154EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWSSVRWNGGIIGYADSVKGRFTISRDNAKNSLYLQMNSLRPEDTALYYCAKDIGDVLTGYYGEYGMDVWGQGTTVTVSS; SEQ ID NO: 155GGA TTC ACC TTT GAT GAT TAT GGC; SEQ ID NO: 156 G F T F D D Y A;SEQ ID NO: 157 GTT AGG TGG AAT GGT GGT ATT ATA; SEQ ID NO: 158V R W N G G I I; SEQ ID NO: 159GCA AAA GAT ATA GGC GAT GTT TTG ACT GGT TAT TAT GGA GAA TAC GGT ATG GAC GTC;SEQ ID NO: 160 A K D I G D V L T G Y Y G E Y G M D V; SEQ ID NO: 161GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTGGGAGACAGAGTCACCATCGCTTGCCGGGCAAGTCAGAGCATTACCACCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGTAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACATTTCCCCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA; SEQ ID NO: 162DIQMTQSPSSLSASVGDRVTIACRASQSITTYLNWYQQKPGKAPKLLIYAASSLQSGVFSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYISPYTFGQGTKLEIK; SEQ ID NO: 163 CAG AGC ATT ACC ACC TAT;SEQ ID NO: 164 Q S I T T Y; SEQ ID NO: 165 GCT GCA TCC; SEQ ID NO: 166A A S; SEQ ID NO: 167 CAA CAG AGT TAC ATT TCC CCG TAG ACT;SEQ ID NO: 168 Q Q S Y I S P Y T; SEQ ID NO: 169CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAAGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAATTATGGCATACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCGATTATATTATATGATGGAAGTAATCAACACTATGCAGATTCCGTGAAGGGCCGATTCACCATTTCCAGAGACAATTCCAAAAACACGCTGTATCTTCAAATGAACAACCTGAGAGCTGAGGACACGGCCGTTTATTACTGTGCGAGAGATCTTGATCTTTGGAGTGGTTATTATACAAACGGGGACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA;SEQ ID NO: 170QVQLVESGGGVVQPGKSLRLSCAASGFTFSNYGIHWVRQAPGKGLEWVAIILYDGSNQHYADSVKGRFTISRDNSKNTLYLQMNNLRAEDTAVYYCARDLDLWSGYYTNGDGMDVWGQGTTVTVSS; SEQ ID NO: 171GGA TTC ACC TTC AGT AAT TAT GGC; SEQ ID NO: 172 G F T F S N Y G;SEQ ID NO: 173 ATA TTA TAT GAT GGA AGT AAT CAA; SEQ ID NO: 174I L Y D G S N Q; SEQ ID NO: 175GCG AGA GAT CTT GAT CTT TGG AGT GGT TAT TAT ACA AAC GGG GAC GGT ATG GAC GTC;SEQ ID NO: 175 A R D L D L W S G Y Y T N G D G M D V;

The amino acid and nucleotide sequences of heavy and light chainimmunoglobulins, including constant domains, of antigen-binding proteinsof the present invention are set forth below:

H4H14699P2 Heavy chain DNA (SEQ ID NO: 179)GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTGCAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCGGCCTCTGGATTCACCTTTGATGATTATGCCATACACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATCAGTTGGAATAGTGATATCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCGTCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAATAGTCTGAGAACTGAGGACACGGCCTTGTATTACTGTGCAAAAGGATATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 180)EVQLVESGGGVPQPGRSLRLSCAASGFTFDDYAIHWVRQAPGKGLEWVSVISWNSDIIGYADSVKGRFTVSRDNAKNSLYLQMNSLRTEDTALYYCAKGYNWNFFDYWGQGTLVTVSSASTKGPSVFPLALCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVNSKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPDLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 181)GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTATCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGCAGCAAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 182)EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYNAANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14700P2Heavy chain DNA (SEQ ID NO: 183)GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAACTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATTAGTTGGAATAGTGATGTCATAGCCTATTCGGACTCTGTGAAGGGCCGCTTCACCATTTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGGGAACTGAGGACACGGCCTTATATTACTGTGCAAAAGGCCATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 184)EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQTPGKGLEWVSVISWNSDVIAYSDSVKGRFTISRDNAKNSLYLQMNSLGTEDTALYYCAKGHNWNFFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVNSKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSVSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 185)GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGAGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGTAGCCAACAGGGCCACAGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCGGCCTAGAGCCTGAAGATTTTGCAGTTTATTTCTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 186)EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYNVANRATDIPARFSGSGSGTDFTLTISGLEPEDFAVYFCQQRSNWPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14706P2Heavy chain DNA (SEQ ID NO: 187)GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTTGATGATTATGCCATACACTGGGTCCGGCAATCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGTTATCAGTTGGAATAGTGATGTCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAGATGAATAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAAGGATATAACTGGAACTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 188)EVQLVESGGGLVQPGRSLRLSCTASGFTFDDYAIHWVRQSPGKGLEWVSVISWNSDVIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKGYNWNFFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVNSKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKRRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 189)GAAATTGTGTTGACGCAGTCTCCAGCCACCCTGTCTTTATCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATAATGCAGCAAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 190)EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYNAANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14708P2Heavy chain DNA (SEQ ID NO: 191)GAAGTGCAGCTGGTGGAGTCTGGGGGAGACTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAATGGGTCTCAGTTATTAGTTGGAATAGTGATGTCATAGCCTATTCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGAACTGAGGACACGGCCTTATATTACTGTACAAAAGGCCATAAGTGGAGCTTCTTTGACTATTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 192)EVQLVESGGDLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSVISWNSDVIAYSDSVKGRFTISRDNAKNSLYLQMNSLRTEDTALYYCTKGHKWSFFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSLGYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVNSKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 193)GAAATTGTGTTGACACAGTCTCCAGCCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTATTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGACTCCTCATCTTTAATGTAGCCAACAGGGCCACTGACATCCCAGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATCAGCAGCCTAGAGCCTGAAGATTTTGCAGTTTATTACTGTCAGCAGCGTAGCAACTGGCCTCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 194)EIVLTQSPATLSLSPGERATLSCRASQSISSYLAWYQQKPGQAPRLLIFNVANRATDIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14709P(SEQ ID NO: 195)GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTTCAGCCTGGGGGGTCCCTGAGACTCTCCTGCGCAGCCTCTGGATTCACCTTTAGCGACTATGCCATGAGCTGGGTCCGCCAGGCTCCGGGGAAGGGGCTGGAGTGGGTCTCAGGTATTAGTGGAAATGGTGGTGACACATACTACGGAGACTTCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGGCGAGGACACGGCCGCATATTTCTGTGTGATAGATCTTGACTATTGGGGTCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGA Heavy chain polypeptide(SEQ ID NO: 196)EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYAMSWVRQAPGKGLEWVSGISGNGGDTYYGDFVKGRFTISRDNSKNTLYLQMNSLRGEDTAAYFCVIDLDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVDLSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 197)GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGAAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAACAGAAACCAGGAAAAGCCCCTAGGCTCCTGATCTATAAGGCGTCTATTTTAGGAGATGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCTACTTATTACTGCCACCAGTATAATAGTTATTTGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 198)DIQMTQSPSTLSASEGDRVTITCRASQSISSWLAWYQQKPGKAPRLLIYKASILGDGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCHQYNSYLWTFGQGTKVEIKRTVAAPSVFIFPPSDELQKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14728PHeavy chain DNA (SEQ ID NO: 199)CAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTGCTGATTACTATTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGATCCATCTATTATACTGGGAGTACTTACTACAACCCGTCCCTCAAGAGTCGACTTACCATATCAATAGACACGTCTGAGAACCAGTTCTCTTTGAAACTGACCTCTCTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGCGAGGAGGCTAACTGGGGATCCCACTTTGACTCCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGCCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 200)QVQLQESGPGLVKPSQTLSLTCTVSGGSISSADYYSWSIRQHPGKGLEWIGSIYYTGSTYYNPSLKSRLTISIDTSENQFSLKLTSLTAADTAVYYCASEEANSGSHFDSWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTEPVTCVVVDVSQEDPEVQRNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 201)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTGACAACTTTTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCATCTTACTACTGTCAACATAGTCACAGTGCCCATCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 202)DIQMTQSPSSLSASVGDRVTITCRASQSIDNFLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFASYYCQHSHSAHPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14731PHeavy chain DNA (SEQ ID NO: 203)CAGCTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCGGAGACCCTGTCCCTCACCTGCACTGTCTCTGGTGGCTCCATCAGCAGTAGTAATTACTACTGGGGCTGGATCCGCCAGCCCCCAGGGAAGAGACTGGAGTGGATTGGGAGTATCTATTATAGTGGGAGCACCTACTACAACCCGTCCCTCAAGACTCGAGTCACCATATCCGTAGACACGTCCAAGAATCAGTTCTCCCTGAAGCTGACCTCTGTGACCGCCGCAGACACGGCTGTGTATTACTGTGCGAGAGAGGAAGCAGCAGCTTTGACGCACTTTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 204)QLQLQESGPGLVKPSETLSLTCTVSGGSISSSNYYWGWIRQPPGKRLEWIGSIYYSGSTYYNPSLKTRVTISVDTSKNQFSLKLTSVTAADTAVYYCAREEAAALTHFDFWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKWRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 205)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAACTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTGCTGCATCCAGTTTACAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACATAGTCACAGTTCCCATCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 206)DIQMTQSPSSLSASVGDRVTITCRASQSISNYLNWYQQKPGKAPKLLIFAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHSHSSHPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14732P2(SEQ ID NO: 207)GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAATTGGGCTGGTTATAACATAGACTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAAGATATGCGTGGATTCAGTTATGGTTTCCCCTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGAHeavy chain polypeptide (SEQ ID NO: 208)EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSGINWAGYNIDYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDMRGFSYGFPFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPEPEQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQEPNNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 209)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCGTCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG Light chain polypeptide (SEQ ID NO: 210)DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H14H14734P2Heavy chain DNA (SEQ ID NO: 211)GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTAAAGCCGGGGGGGTCCCTTAGACTCTCCTGTGCAGCCTCTGGATTTATTTTCAGTAACGCCTGGATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGACTGGCGTGGGTTGGCCGTATTAAAACCGAAACTGATGGTGGGACAACAGACTACGCTGCACCCGTAAAAGGCAGATTCACCATCTCAAGAGATGACTCAAAAAACACTCTGTATCTGCAAATGAACAGCGTGAAAACCGAGGACACAGCCGTGTATTACTGTACAGGGGGATACAGCTATGGTGACGATAGCAGCAGCTGGAACGAGGGCTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGA Heavy chain polypeptide (SEQ ID NO: 212)EVQLVESGGGLVKPGGSLRLSCAASGFIFSNAWMNWVRQAPGKGLAWVGRIKTETDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSVKTEDTAVYYCTGGYSYGDDSSSWNEGYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKLight chain DNA (SEQ ID NO: 213)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCGTCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG Light chain polypeptide (SEQ ID NO: 214)DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14757PHeavy chain DNA (SEQ ID NO: 215)GAAGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTCGTTGGAATGGTGGTAGTATAGGCTATGTGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAAGTCCCTGCATCTGCAAATGAACAGTCTAAAAACTGAGGACACGGCCTTGTATTACTGTGCAAAAGATATAGGCGATATTTTGACTGGTTTTTATGGAGAATACGGAATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGA Heavy chain polypeptide (SEQ ID NO: 216)EVQLVESGGGLVQPGRSLRLSCTASGFTFDDYAMHWVRQAPGKGLEWVSGIRWNGGSIGYVDSVKGRFTISRDNAKKSLHLQMNSLKTEDTALYYCAKDIGDILTGFYGEYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 217)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGAAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAAGCAGGGAAAGCCCCTAACCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACATTATCCCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 218)DIQMTQSPSSLSASEGDRVTITCRASQSISSYLNWYQQKAGKAPNLLIYAASSLQSGVPSRFSGSGSGTEYTLTISSLQPEDFATYYCQQSYIIPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLAAPVTKSFNRGEC H4H14758PHeavy chain DNA (SEQ ID NO: 219)GAAGTGCAGCTGGTGGAGTCTGGGGGAGGGTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAAGTGTTAGGTGGAATGGTGGTATTATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACAGTCTGAGACCTGAGGACACGGCCCTCTATTACTGTGCAAAAGATATAGGCGATGTTTTGACTGGTTATTATGGAGAATACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGA Heavy chain polypeptide (SEQ ID NO: 220)EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSSVRWNGGIIGYADSVKGRFTISRDNAKNSLYLQMNSLRPEDTALYYCAKDIGDVLTGYYGEYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 221)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTGGGAGACAGAGTCACCATCGCTTGCCGGGCAAGTCAGAGCATTACCACCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGTAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACATTTCCCCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT Light chain polypeptide (SEQ ID NO: 222)DIQMTQSPSSLSASVGDRVTIACRASQSITTYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYISPYTFGQGTKLEIKRTVAASPVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC H4H14760P2Heavy chain DNA (SEQ ID NO: 223)CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAAGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAATTATGGCATACACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCGATTATATTATATGATGGAAGTAATCAACACTATGCAGATTCCGTGAAGGGCCGATTCACCATTTCCAGAGACAATTCCAAAAACACGCTGTATCTTCAAATGAACAACCTGAGAGCTGAGGACACGGCCGTTTATTACTGTGCGAGAGATCTTGATCTTTGGAGTGGTTATTATACAAACGGGGACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCACCCTGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTCACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGTCCCTCTCCCTGTCTCTGGGTAAATGA Heavy chain polypeptide (SEQ ID NO: 224)QVLQVESGGGVVQPGKSLRLSCAASGFTFSNYGIHWVRQAPGKGLEWVAIILYDGSNQHYADSVKGRFTISRDNSKNTLYLQMNNLRAEDTAVYYCARDLDLWSGYYTNGDGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTRYVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK Light chain DNA(SEQ ID NO: 225)GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGAGCATTAGCAGCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCGTCAAGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGAGTTACAGTACCCCTCCGATCACCTTCGGCCAAGGGACACGACTGGAGATTAAAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG Light chain polypeptide (SEQ ID NO: 226)DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIKTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

Example 2: Bioassay with HEK293/D9(NFκB-luciferase)/hIL-36R andHEK293/NFκB-luciferase/mfIL-36R Cells

IL-36 receptor (IL-36R) is a single-pass membrane receptor for a subsetof members of the IL-1 family of cytokines, IL-36α, IL-36β, and IL-36γ,and upon binding to these ligands, there is recruitment of itsco-receptor, the IL-1R accessory protein (IL-1RAcP), which induces asignaling cascade that involves NFκB and mitogen-activated kinasepathways (Sims et al, 2010). A bioassay was developed to detect thetranscriptional activation by NFκB via IL-36R activation using reportercell lines that stably express either full-length human IL-36R (hIL-36R;amino acids 1 through 575 of accession number NP_003845.2) or Macacafascicularis IL-36R (MfIL-36R) along with a luciferase reporter [NFκBresponse element (5×)-luciferase-IRES-GFP] in HEK293 cells. IL-1RAcP isendogenously expressed in the HEK293 cell line. The resulting stablecell lines, referred to as HEK293/NFκB-luc/hIL-36R andHEK293/NFκB-luc/MfIL-36R, was isolated and maintained in DMEM containing10% FBS, NEAA, penicillin/streptomycin/glutamine, and 500 μg/mL G418.

For the bioassay, cells were seeded into 96-well assay plates at 10,000cells/well in OPTIMEM supplemented with 0.1% FBS and then incubated at37° C. in 5% CO₂ overnight. The next day, to determine the dose responseof ligands, human IL-36α (hIL-36α; R&D Systems, #6995/IL), human IL-36β(hIL-36β; R&D Systems, #6334-IL), or human IL-36γ (hIL-36γ; R&D Systems,#6835-IL) were serially diluted at 1:3 (from 10 nM to 0.0002 nM) andadded to cells. A control containing dilution buffer but no IL-36 ligandwas also added to one sample of cells. To measure inhibition, antibodieswere serially diluted at 1:3 (from 100 nM to 0.002 nM) plus a controlsample containing no antibody and pre-incubated with the cells followedby addition of constant concentrations of hIL-36α, hIL-36β, or hIL-36γ.For testing with HEK293/NFκB-luc/hIL-36R cells, 20 pM of hIL-36α, 15 pMof hIL-36β, or 10 pM of hIL-36γ was used as a constant concentration andfor testing with HEK293/NFκB-luc/mfIL-36R cells, 500 pM of hIL-36α, 600pM of hIL-36β, or 300 pM of hIL-36γ was used as a constantconcentration. After 5.5 hours of incubation at 37° C. in 5% CO₂, OneGloreagent (Promega, # E6051) was added to the samples and luciferaseactivity was then measured using a Victor X (Perkin Elmer) plate reader.

The results were analyzed using nonlinear regression (4-parameterlogistics) with Prism 6 software (GraphPad) to obtain EC₅₀ and IC₅₀values. To determine the maximum inhibition, the range between themaximum and minimum RLU values for each antibody was calculated as apercentage of the RLU range between no IL-36 ligand and the constantamount of IL-36 ligand used per assay.

As shown in Table 2-1, 9 out of 12 anti-IL-36R antibodies of theinvention tested completely blocked the stimulation ofHEK293/NFkB-luc/hIL-36R cells by 20 pM hIL-36α with IC₅₀ values rangingfrom 100 pM to 970 pM. One of the IL-36R antibodies tested demonstratedpartial blockade of hIL-36α stimulation of HEK293/NFkB-luc/hIL-36R cellswith a maximum percent blockade of 22%. One of the IL-36R antibodiestested demonstrated weak blockade of hIL-36α stimulation ofHEK293/NFkB-luc/hIL-36R cells with a maximum percent blockade of 61%,while another of the anti-IL-36R antibodies tested did not demonstratedany inhibition of hIL-36α stimulation. Six out of 12 anti-IL-36Rantibodies of the invention tested completely blocked the stimulation ofHEK293/NFkB-luc/hIL-36R cells by 15 pM hIL-36β with IC₅₀ values rangingfrom 120 pM to 1.3 nM. One of the IL-36R antibodies tested demonstratedweak blockade of hIL-36β stimulation of HEK293/NFkB-luc/hIL-36R cellswith a maximum percent blockade of 69% and 5 anti-IL-36R antibodiestested did not demonstrate measurable inhibition of hIL-36β stimulation.Six out of 12 anti-IL-36R antibodies of the invention tested completelyblocked the stimulation of HEK293/NFkB-luc/hIL-36R cells by 10 pMhIL-36γ with IC₅₀ values ranging from 120 pM to 1.2 nM. Four anti-IL-36Rantibodies of the invention tested demonstrated partial blockade ofhIL-36γ stimulation with maximum percent blockade ranging from 24 to87%. One anti-IL-36R antibody of the invention tested showed weakblockade of hIL-36γ stimulation with maximum percent blockade of 69%,and one anti-IL36R antibody of the invention did not demonstrateinhibition of hIL-36γ stimulation. The isotype control antibody testeddid not demonstrate inhibition of IL-36 ligand stimulation of theHEK293/NFkB-luc/hIL-36R cells. As shown in Table 2-1, hIL-36α, hIL-36β,and hIL-36γ activated HEK293/NFkB-luc/hIL-36R cells with EC₅₀ values of12 pM, 14 pM, and 8.4 pM respectively.

As shown in Table 2-2, six out of 12 anti-IL-36R antibodies of theinvention tested completely or nearly completely blocked the stimulationof HEK293/NFkB-luc/MfIL-36R cells by 500 pM hIL-36α with IC₅₀ valuesranging from 60 pM to 3.1 nM. Two anti-IL-36R antibodies of theinvention tested demonstrated weak blockade of hIL-36α stimulation ofHEK293/NFkB-luc/MfIL-36R cells with maximum percent blockade of 29 and47%, while 4 anti-IL-36R antibodies did not show inhibition of hIL-36αstimulation of this cell line. Six out of 12 anti-IL-36R antibodies ofthe invention tested completely or nearly completely blocked thestimulation of HEK293/NFkB-luc/MfIL-36R cells by 600 pM hIL-36β withIC₅₀ values ranging from 120 pM to 7.1 nM. Three anti-IL-36R antibodiesof the invention tested demonstrated weak blockade of hIL-36βstimulation of HEK293/NFkB-luc/MfIL-36R cells with maximum percentblockade ranging from 36 to 48%, while three anti-IL-36R antibodies ofthe invention did not show inhibition of hIL-36β stimulation of thiscell line. Six out of anti-IL-36R antibodies of the invention testedcompletely or nearly completely blocked the stimulation ofHEK293/NFkB-luc/MfIL-36R cells by 300 pM hIL-36γ with IC₅₀ valuesranging from 85 pM to 5.4 nM. Three anti-IL-36R antibodies of theinvention tested showed weak blockade of hIL-36γ stimulation ofHEK293/NFkB-luc/MfIL-36R cells with maximum percent blockade rangingfrom 25 to 43%, while three anti-IL-36R antibodies of the invention didnot show inhibition of hIL-36γ stimulation of this cell line. Theisotype control antibody tested did not demonstrate inhibition of IL-36ligand stimulation of the HEK293/NFkB-luc/MfIL-36R cells. As shown inTable 2-1, hIL-36α, hIL-36β, and hIL-36γ activatedHEK293/NFkB-luc/MfIL-36R cells with EC₅₀ values of 170 pM, 270 pM, and62 pM respectively.

TABLE 2-1 Anti-IL-36R antibody inhibition of stimulation ofHEK293/NFκB-luc/hIL-36R cells by hIL-36 ligands. Ligand hIL-36α hIL-36βhIL-36γ EC₅₀ 1.2E−11M 1.4E−11M 8.4E−12M Constant 20 pM 15 pM 10 pM MaxMax Max Inhibition Inhibition Inhibition Antibodies IC₅₀ [M] (%) IC₅₀[M] (%) IC₅₀ [M] (%) H4H14699P2 1.3E−10 100 1.3E−10 100 1.4E−10 99H4H14700P2 1.9E−10 101 2.0E−10 100 1.2E−10 100  H4H14706P2 1.0E−10 1011.2E−10 101 1.2E−10 99 H4H14708P2 1.3E−10 101 2.0E−10 100 1.6E−10 99H4H14709P 1.4E−10 22 No No 1.3E−10 24 (partial) inhibition inhibition(partial) H4H14728P 9.7E−10 97 1.3E−09 99 1.2E−09 99 H4H14731P 7.8E−1099 9.4E−10 99 7.3E−10 99 H4H14732P2 >1.0E−08  61 >1.0E−08  69 >1.0E−08 69 H4H14734P2 No No No No No No inhibition inhibition inhibitioninhibition inhibition inhibition H4H14757P 1.8E−10 101 No No 1.3E−10 87inhibition inhibition (partial) H4H14758P 1.2E−10 100 No No 1.6E−10 57inhibition inhibition (partial) H4H14760P2 4.9E−10 99 No No 7.0E−10 49inhibition inhibition (partial) Isotype No No No No No No controlinhibition inhibition inhibition inhibition inhibition inhibitionantibody

TABLE 2-2 Anti-IL-36R antibody inhibition of stimulation ofHEK293/NFκB-luc/MfIL-36R cells by hIL-36 ligands. Ligand hIL-36α hIL-36βhIL-36γ EC₅₀ 1.7E−10M 2.7E−10M 6.2E−11M Constant 500 pM 600 pM 300 pMMax Max Max Inhibition Inhibition Inhibition Antibodies IC₅₀ [M] (%)IC₅₀ [M] (%) IC₅₀ [M] (%) H4H14699P2 8.4E−11 97 1.9E−10 98 2.0E−10 97H4H14700P2 1.2E−10 99 1.8E−10 99 1.8E−10 99 H4H14706P2 6.0E−11 100 1.2E−10 100  8.5E−11 100  H4H14708P2 8.9E−11 99 1.2E−10 100  1.2E−10100  H4H14709P No No No No No No inhibition inhibition inhibitioninhibition inhibition inhibition H4H14728P 1.3E−09 93 1.5E−09 95 2.0E−0993 H4H14731P 3.1E−09 84 7.1E−09 78 5.4E−09 75 H4H14732P2 >1.0E−07 47 >1.0E−07  43 >1.0E−07  36 H4H14734P2 No No No No No No inhibitioninhibition inhibition inhibition inhibition inhibitionH4H14757P >1.0E−07  29 >1.0E−07  36 >1.0E−07  25 H4H14758P No No No NoNo No inhibition inhibition inhibition inhibition inhibition inhibitionH4H14760P2 No No >1.0E−07  48 >1.0E−07  43 inhibition inhibition IsotypeNo No No No No No control inhibition inhibition inhibition inhibitioninhibition inhibition antibody

Example 3: IL-36R Octet Cross-Competition

Binding competition between a panel of different anti-IL-36R antibodieswas determined using a real time, label-free bio-layer interferometryassay on an Octet® HTX biosensor (ForteBio, A Division of Pall LifeSciences). The entire experiment was performed at 25° C. in 0.01M HEPESpH 7.4, 0.15 M NaCl, 3 mM EDTA, 0.05% v/v Surfactant Tween-20, 0.002%NaN₃ and 1 mg/mL BSA (HBS-ET kinetics buffer) with the plate shaking atthe speed of 1000 rpm. To assess whether two antibodies are able tocompete with one another for binding to their respective epitopes on therecombinant human IL-36R extracellular domain expressed with aC-terminal myc-myc-hexahistidine tag (hIL-36R-MMH: mROR1 signal sequence(M1-A29)-human IL36R(D20-Y337)-mycmycHis₆), around 0.3 nM of hIL-36R-MMHwas first captured onto anti-His antibody coated Octet biosensors(Fortebio Inc, #18-5079) by submerging the biosensors for 3 minutes intowells containing 30 μg/mL of hIL-36R-MMH. The antigen-capturedbiosensors were then saturated with a first anti-IL-36R antibody(subsequently referred to as mAb-1) by submerging into wells containing50 μg/mL solution of mAb-1 for 4 minutes. The biosensors were thensubsequently submerged into wells containing a 50 μg/mL solution of asecond anti-IL-36R antibody (subsequently referred to as mAb-2) for 3minutes. The biosensors were washed in HBS-ET kinetics buffer in betweenevery step of the experiment. The real-time binding response wasmonitored during the entire course of the experiment and the maximumbinding response for all the steps was recorded. The response of mAb-2binding to hIL-36R-MMH pre-complexed with mAb-1 was compared andcompetitive/non-competitive behavior of different anti-IL-36R antibodieswas determined as shown in Table 3-1.

TABLE 3-1 Cross-competition of anti-IL-36R antibodies for binding tohuman IL-36R-MMH First antibody (mAb-1) Second antibody binding tocaptured (mAb-2) shown to hIL-36R-MMH compete with mAb-1 H4H14699P2H4H14700P2 H4H14706P2 H4H14708P2 H4H14732P2 H4H14700P2 H4H14699P2H4H14706P2 H4H14708P2 H4H14732P2 H4H14706P2 H4H14699P2 H4H14700P2H4H14708P2 H4H14732P2 H4H14708P2 H4H14699P2 H4H14700P2 H4H14706P2H4H14732P2 H4H14732P2 H4H14699P2 H4H14700P2 H4H14706P2 H4H14708P2H4H14757P H4H14758P H4H14760P2 H4H14758P H4H14757P H4H14760P2 H4H14760P2H4H14757P H4H14758P H4H14728P H4H14731P H4H14731P H4H14728P H4H14709PH4H14734P2 H4H14734P2 H4H14709P

Example 4: Antibody Binding Kinetics

Equilibrium dissociation constants (K_(D) values) for IL-36R binding topurified anti-IL-36R antibodies were determined using a real-timesurface plasmon resonance biosensor using a Biacore 4000 instrument. TheBiacore sensor surface was first derivatized by amine coupling with amonoclonal mouse anti-human Fc antibody (GE, # BR-1008-39) to captureanti-IL-36R monoclonal antibodies. All binding studies were performed in0.01 M Hepes pH 7.4, 0.15 M NaCl, 3 mM EDTA, and 0.05% v/v SurfactantTween-20 (HBS-ET running buffer) at 25° C. and 37° C. Differentconcentrations of IL-36R reagents, human IL-36R extracellular domainexpressed with a C-terminal myc-myc-hexahistidine tag (hIL-36R-MMH),Macaca fascicularis IL-36R extracellular domain expressed with aC-terminal myc-myc-hexahistidine tag (mfIL-36R-MMH: mROR1 signalsequence (M1-A29). Macaca fascicularis IL36R_ecto domain(D20-A336).mycmycHis6), human IL-36R extracellular domain expressed witha C-terminal mouse IgG2a Fc tag (hIL-36R-mFc: mROR1 signal sequence(M1-A29)-human IL36R (D20-Y337)-mouse IgG2aFc (E98-K330)) or an in-linefusion protein of human IL-36R extracellular domain and IL1RAcPextracellular domain expressed with mouse IgG2a Fc tag(hIL-36R-Trap-mFc: mROR1 signal sequence (M1-A29)-human IL36R ectodomain (D20-Y337)-human IL1RacP ecto domain(S21-E359)-mouse IgG2aFc) inHBS-ET running buffer (ranging from 100 nM to 3.7 nM, 3-fold dilutions)were injected over the anti-IL-36R antibody captured surface for 4minutes at a flow rate of 30 μL/minute and their dissociation in HBS-ETrunning buffer was monitored for 10 minutes. Kinetic association rateconstant (k_(a)) and dissociation rate constant (k_(d)) were determinedby fitting the real-time sensorgrams to a 1:1 binding model usingScrubber 2.0c curve fitting software. Binding dissociation equilibriumconstants (K_(D)) and dissociative half-lives (t_(1/2)) were calculatedfrom the kinetic rate constants as:

${{K_{D}(M)} = \frac{kd}{ka}},{{{and}\mspace{14mu} {t^{1/2}\left( \min \right)}} = \frac{\ln (2)}{60*{kd}}}$

Binding kinetic parameters for hIL-36R-MMH, mfIL-36R-MMH or hIL-36R.mFcbinding to different anti-IL-36R antibodies of the invention at 25° C.and 37° C. are shown in Tables 4-1 through 4-8. At 25° C., hIL-36R-MMHbound to all of the anti-IL-36R antibodies of the invention with K_(D)values ranging from 2.18 nM to 13.9 nM, as shown in Table 4-1. At 37°C., hIL-36R-MMH bound to all of the anti-IL-36R antibodies of theinvention with K_(D) values ranging from 4.25 nM to 29.5 nM, as shown inTable 4-2. At 25° C., mfIL-36R-MMH bound to 9 of the 12 anti-IL-36Rantibodies of the invention with K_(D) values ranging from 7.87 nM to34.4 nM, as shown in Table 4-3. At 37° C., mfIL-36R-MMH bound to 9 ofthe 12 anti-IL-36R antibodies of the invention with K_(D) values rangingfrom 14.4 nM to 58.2 nM, as shown in Table 4-4. At 25° C., hIL-36R-mFcbound to 11 of the 12 anti-IL-36R antibodies of the invention with K_(D)values ranging from 173 pM to 5.79 nM, as shown in Table 4-5. Oneanti-IL-36R antibody of the invention demonstrated inconclusive bindingto hIL-36R-mFc under the experimental conditions at 25° C. At 37° C.,hIL-36R-mFc bound to all of the anti-IL-36R antibodies of the inventionwith K_(D) values ranging from 205 pM to 28.7 nM, as shown in Table 4-6.At 25° C., hIL-36R-Trap-mFc bound to all of the anti-IL-36R antibodiesof the invention with K_(D) values ranging from 212 pM to 14 nM, asshown in Table 4-7. At 37° C., hIL-36R-Trap-mFc bound to all of theanti-IL-36R antibodies of the invention with K_(D) values ranging from264 pM to 40.9 nM, as shown in Table 4-8.

TABLE 4-1 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R-MMH at 25° C. mAb 100 nM Capture hIL-36R-MMH k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 198 ± 0.6 74 1.65E+05 1.68E−03 1.02E−08 7 H4H14700P2 159 ±0.3 67 1.20E+05 5.79E−04 4.82E−09 20 H4H14706P2 199 ± 0.5 89 1.21E+054.95E−04 4.08E−09 23 H4H14708P2 209 ± 0.9 76 9.14E+04 6.23E−04 6.82E−0919 H4H14709P 156 ± 0.2 64 7.23E+04 2.96E−04 4.09E−09 39 H4H14728P 175 ±0.6 69 9.83E+04 7.73E−04 7.87E−09 15 H4H14731P 204 ± 0.6 54 9.22E+043.43E−04 3.72E−09 34 H4H14732P2 197 ± 0.3 26 4.10E+04 5.69E−04 1.39E−0820 H4H14734P2 174 ± 0.8 22 3.32E+04 4.04E−04 1.22E−08 29 H4H14757P 180 ±0.9 94 1.82E+05 3.96E−04 2.18E−09 29 H4H14758P 177 ± 0.7 87 1.21E+059.23E−04 7.63E−09 13 H4H14760P2 180 ± 0.5 61 6.79E+04 4.15E−04 6.11E−0928

TABLE 4-2 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R-MMH at 37° C. mAb 100 nM Capture hIL-36R-MMH k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 257 ± 1.2 89 1.42E+05 4.18E−03 2.95E−08 2.8 H4H14700P2 218 ±0.7 89 1.76E+05 1.84E−03 1.05E−08 6 H4H14706P2 266 ± 1   113 1.50E+051.30E−03 8.65E−09 9 H4H14708P2 280 ± 2.5 106 1.36E+05 1.86E−03 1.36E−086 H4H14709P 218 ± 0.9 106 1.31E+05 5.54E−04 4.25E−09 21 H4H14728P 242 ±0.7 93 1.36E+05 2.93E−03 2.15E−08 4 H4H14731P 262 ± 0.7 81 1.37E+059.71E−04 7.11E−09 12 H4H14732P2 272 ± 0.8 34 4.21E+04 1.16E−03 2.76E−0810 H4H14734P2 248 ± 0.8 25 5.39E+04 7.49E−04 1.39E−08 15 H4H14757P 262 ±1.0 129 2.10E+05 1.09E−03 5.18E−09 11 H4H14758P 247 ± 1.2 111 1.58E+052.37E−03 1.50E−08 5 H4H14760P2 252 ± 0.8 83 9.06E+04 2.08E−03 2.30E−08 6

TABLE 4-3 Binding Kinetics parameters of anti-IL-36R antibodies bindingto mfIL-36R-MMH at 25° C. mAb 100 nM Capture mfIL-36R-MMH k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 198 ± 0.4 53 6.38E+04 1.83E−03 2.87E−08 6 H4H14700P2 158 ±0.3 48 6.02E+04 6.35E−04 1.06E−08 18 H4H14706P2 199 ± 0.4 65 6.72E+045.52E−04 8.21E−09 21 H4H14708P2 209 ± 0.9 51 4.50E+04 5.79E−04 1.29E−0820 H4H14709P 156 ± 0.3 34 2.80E+04 4.01E−04 1.43E−08 29 H4H14728P 175 ±0.6 51 5.15E+04 4.06E−04 7.87E−09 28 H4H14731P 203 ± 0.4 33 5.98E+049.51E−04 1.59E−08 12 H4H14732P2 197 ± 0.4 12 1.88E+04 6.49E−04 3.44E−0818 H4H14734P2 175 ± 0.6 12 2.60E+04 2.54E−04 9.78E−09 46 H4H14757P 183 ±0.7 1 NB* NB* NB* NB* H4H14758P 179 ± 0.6 −1 NB* NB* NB* NB* H4H14760P2181 ± 0.4 0 NB* NB* NB* NB* *NB indicates that under the experimentalconditions, mfIL-36R-MMH reagent did not bind to the capturedanti-IL-36R antibody

TABLE 4-4 Binding Kinetics parameters of anti-IL-36R antibodies bindingto mflL-36R-MMH at 37° C. mAb 100 nM Capture mfIL-36R-MMH k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 257 ± 0.4 59 7.80E+04 4.54E−03 5.82E−08 2.5 H4H14700P2 218 ±0.7 67 7.14E+04 1.98E−03 2.78E−08 6 H4H14706P2 266 ± 0.4 84 7.47E+041.42E−03 1.90E−08 8 H4H14708P2 279 ± 2.8 75 5.96E+04 1.81E−03 3.04E−08 6H4H14709P 220 ± 1.4 66 4.91E+04 8.71E−04 1.77E−08 13 H4H14728P 243 ± 0.777 6.48E+04 1.34E−03 2.07E−08 9 H4H14731P 261 ± 0.3 41 6.68E+04 3.22E−034.82E−08 4 H4H14732P2 273 ± 0.8 17 3.19E+04 1.64E−03 5.15E−08 7H4H14734P2 248 ± 0.6 12 3.61E+04 5.21E−04 1.44E−08 22 H4H14757P 264 ±1.5 4 NB* NB* NB* NB* H4H14758P 248 ± 0.9 −1 NB* NB* NB* NB* H4H14760P2253 ± 0.9 2 NB* NB* NB* NB* *NB indicates that under the experimentalconditions, mfIL-36R-MMH reagent did not bind to the capturedanti-IL-36R antibody

TABLE 4-5 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R-mFc at 25° C. mAb 100 nM Capture hIL-36R-mFc k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 197 ± 0.8 150 5.64E+05 2.03E−04 3.59E−10 57 H4H14700P2 158 ±0.3 128 5.17E+05 1.20E−04 2.33E−10 96 H4H14706P2 197 ± 1.7 163 5.75E+051.32E−04 2.30E−10 87 H4H14708P2 207 ± 2.3 146 4.03E+05 1.28E−04 3.17E−1090 H4H14709P 155 ± 0.7 142 2.57E+05 8.62E−05 3.35E−10 134 H4H14728P 174± 0.5 7 IC* IC* IC* IC* H4H14731P 204 ± 0.3 10 5.58E+04 3.23E−045.79E−09 36 H4H14732P2 197 ± 0.6 145 5.70E+05 7.97E−04 1.40E−09 14H4H14734P2 174 ± 0.5 77 6.22E+04 1.01E−04 1.63E−09 114 H4H14757P 182 ±1.4 167 5.75E+05 9.97E−05 1.73E−10 116 H4H14758P 179 ± 0.8 161 5.26E+051.63E−04 3.09E−10 71 H4H14760P2 181 ± 0.8 121 2.07E+05 1.09E−04 5.26E−10106 *IC indicates that under the experimental conditions, hIL-36R.mFcbinding is inconclusive

TABLE 4-6 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R-mFc at 37° C. mAb 100 nM Capture hIL-36R-mFc k_(a) k_(d)K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 258 ± 0.7 186 5.94E+05 4.56E−04 7.67E−10 25 H4H14700P2 218 ±0.5 174 5.35E+05 2.09E−04 3.90E−10 55 H4H14706P2 266 ± 0.6 207 5.93E+052.66E−04 4.49E−10 43 H4H14708P2 280 ± 2.5 203 4.60E+05 2.00E−04 4.35E−1058 H4H14709P 218 ± 1.2 211 5.43E+05 1.12E−04 2.05E−10 104 H4H14728P 243± 1.0 11 1.62E+04 4.65E−04 2.87E−08 25 H4H14731P 261 ± 0.5 12 6.99E+044.50E−04 6.43E−09 26 H4H14732P2 273 ± 1.2 195 6.35E+05 1.27E−03 2.00E−099 H4H14734P2 247 ± 1.0 96 5.27E+04 1.22E−04 2.31E−09 95 H4H14757P 264 ±2.0 235 6.16E+05 1.50E−04 2.43E−10 77 H4H14758P 248 ± 0.7 210 5.54E+052.86E−04 5.17E−10 40 H4H14760P2 254 ± 0.9 173 2.17E+05 2.22E−04 1.02E−0952

TABLE 4-7 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R-Trap-mFc at 25° C. mAb 100 nM Capture hIL-36R-Trap-mFc k_(a)k_(d) K_(D) t½ Antibody Level (RU) Bound (RU) (1/Ms) (1/s) (M) (min)H4H14699P2 196 ± 0.7 188 4.98E+05 2.28E−04 4.58E−10 51 H4H14700P2 156 ±0.7 157 4.42E+05 1.25E−04 2.83E−10 92 H4H14706P2 195 ± 0.6 205 4.77E+051.10E−04 2.32E−10 105 H4H14708P2 205 ± 2.1 172 3.61E+05 1.29E−043.57E−10 90 H4H14709P 155 ± 0.4 173 2.00E+05 8.07E−05 4.04E−10 143H4H14728P 175 ± 0.5 63 4.03E+04 5.65E−04 1.40E−08 20 H4H14731P 203 ± 0.560 4.52E+04 2.01E−04 4.45E−09 57 H4H14732P2 197 ± 0.4 161 4.62E+051.36E−03 2.95E−09 8 H4H14734P2 174 ± 0.5 85 4.89E+04 1.10E−04 2.24E−09105 H4H14757P 181 ± 0.5 202 4.85E+05 1.03E−04 2.12E−10 113 H4H14758P 179± 0.6 197 4.36E+05 1.80E−04 4.13E−10 64 H4H14760P2 181 ± 1.0 1341.58E+05 1.00E−04 6.36E−10 115

TABLE 4-8 Binding Kinetics parameters of anti-IL-36R antibodies bindingto hIL-36R- Trap-mFc at 37° C. 100 nM hIL- mAb 36R-Trap- Capture mFcBound k_(a) k_(d) K_(D) t½ Antibody Level (RU) (RU) (1/Ms) (1/s) (M)(min) H4H14699P2 256 ± 0.8 229 4.84E+05 5.13E−04 1.06E−09 23 H4H14700P2217 ± 0.5 216 4.55E+05 2.53E−04 5.55E−10 46 H4H14706P2 266 ± 0.8 2644.91E+05 2.71E−04 5.51E−10 43 H4H14708P2 280 ± 1   239 3.85E+05 2.53E−046.56E−10 46 H4H14709P 218 ± 1.3 257 4.67E+05 1.23E−04 2.64E−10 94H4H14728P 243 ± 0.5 89 4.85E+04 1.98E−03 4.09E−08 6 H4H14731P 261 ± 0.478 5.26E+04 5.14E−04 9.77E−09 22 H4H14732P2 272 ± 0.7 212 5.01E+051.82E−03 3.63E−09 6 H4H14734P2 248 ± 0.5 99 4.84E+04 1.43E−04 2.96E−0981 H4H14757P 263 ± 1.2 281 5.23E+05 1.96E−04 3.74E−10 59 H4H14758P 248 ±0.6 251 4.68E+05 3.63E−04 7.77E−10 32 H4H14760P2 254 ± 1.1 195 1.66E+052.83E−04 1.70E−09 41

Additional binding experiments were performed to determine the effect ofpH on the rate of dissociation of IL-36R bound to purified anti-IL-36Rantibodies, which was determined using a real-time surface plasmonresonance biosensor using a Biacore T200 instrument. The Biacore sensorsurface was first derivatized by amine coupling with a monoclonal mouseanti-human Fc antibody (GE, # BR-1008-39) to capture anti-IL-36Rantibodies. These Biacore binding studies were performed using tworunning buffers PBS-T, pH7.4 (8.1 mM Na₂HPO₄, 1.9 mM NaH₂PO₄, 3 mM KCl,137 mM NaCl, 0.05% v/v Tween-20, adjusted to pH 7.4) and PBS-T, pH 6.0(6.6 mM Na₂HPO₄, 3.4 mM NaH₂PO₄, 3 mM KCl, 137 mM NaCl, 0.05% v/vTween-20, adjusted to pH6.0). Different concentrations of hIL-36R-MMHand mfIL-36R-MMH prepared in PBS-T, pH7.4 buffer (ranging from 100 nM to11.11 nM, 3-fold dilutions) were injected over the anti-IL-36R antibodycaptured surface for 4 minutes at a flow rate of 50 μL/minute and theirdissociation in two running buffers, PBS-T, pH7.4 and PBS-T, pH 6.0, wasmonitored for 10 minutes. All of these binding kinetics experiments wereperformed at 25° C. and 37° C. Kinetic dissociation constant (k_(d))were determined by fitting the real-time sensorgrams to a 1:1 bindingmodel using Scrubber 2.0c curve fitting software. Binding dissociativehalf-lives (t_(1/2)) were calculated from k_(d) as:

${t^{1/2}\left( \min \right)} = \frac{\ln (2)}{60*{kd}}$

Binding dissociation rate constants for hIL-36R-MMH or mfIL-36R-MMHbinding to different anti-IL-36R antibodies at 25° C. and 37° C. in tworunning buffers PBS-T, pH7.4 and PBS-T, pH 6.0 are shown in Tables 4-9through 4-12.

TABLE 4-9 Binding dissociation rate constant of anti-IL-36R monoclonalantibodies binding to hIL-36R-MMH in two running buffers performed at25° C. PBS-T, pH 7.4 Running Buffer PBS-T, pH 6.0 Running Buffer 100 nM100 nM Human Human mAb IL-36R- mAb IL-36R- Capture MMH Capture MMH LevelBound k_(d) t½ Level Bound k_(d) t½ mAb PID (RU) (RU) (1/s) (min) (RU)(RU) (1/s) (min) H4H14699P2 255 ± 2.7 84 1.93E−03 6 278 ± 0.6 792.93E−03 4 H4H14700P2 330 ± 0.9 128 7.95E−04 15 350 ± 0.9 120 1.72E−03 7H4H14706P2 298 ± 0.9 119 6.16E−04 19 312 ± 2.2 109 9.27E−04 12H4H14708P2 268 ± 2.8 87 7.96E−04 15 283 ± 2.5 79 1.63E−03 7 H4H14709P283 ± 0.8 111 4.20E−04 28 300 ± 1.8 99 8.42E−04 14 H4H14728P 265 ± 1.7103 8.98E−04 13 269 ± 2   96 8.87E−04 13 H4H14731P 282 ± 1.9 65 5.53E−0421 281 ± 0.9 43 6.79E−04 17 H4H14732P2 244 ± 1.5 32 8.65E−04 13 255 ±1.3 28 9.18E−04 13 H4H14734P2 230 ± 1.3 20 7.38E−04 16 240 ± 1.8 187.87E−04 15 H4H14757P 226 ± 0.6 105 5.97E−04 19 235 ± 1.5 98 8.87E−04 13H4H14758P 244 ± 2.5 108 1.12E−03 10 255 ± 1.6 103 1.80E−03 6 H4H14760P2257 ± 1.5 80 5.45E−04 21 266 ± 1.3 69 9.72E−04 12

TABLE 4-10 Binding dissociation rate constant of anti-IL-36R monoclonalantibodies binding to hIL-36R-MMH in two running buffers performed at37° C. PBS-T, pH 7.4 Running Buffer PBS-T, pH 6.0 Running Buffer 100 nM100 nM hIL- hIL- mAb 36R- mAb 36R- Capture MMH Capture MMH Level Boundk_(d) t½ Level Bound k_(d) t½ mAb PID (RU) (RU) (1/s) (min) (RU) (RU)(1/s) (min) H4H14699P2 312 ± 2.9 96 4.15E−03 3  300 ± 14.2 83 5.81E−03 2H4H14700P2 422 ± 5.7 153 1.69E−03 7 435 ± 1.5 144 3.50E−03 3 H4H14706P2367 ± 3.5 140 1.31E−03 9 378 ± 2   132 2.33E−03 5 H4H14708P2 313 ± 6.8105 1.77E−03 7 318 ± 4.6 95 3.37E−03 3 H4H14709P 372 ± 3   159 7.04E−0416 380 ± 2.9 146 1.72E−03 7 H4H14728P 306 ± 1   121 2.96E−03 4 302 ± 1.3114 3.06E−03 4 H4H14731P 272 ± 3.9 91 1.08E−03 11 276 ± 1.6 84 2.01E−036 H4H14732P2 303 ± 3   40 1.10E−03 10 310 ± 2   36 1.23E−03 9 H4H14734P2287 ± 1.4 20 1.21E−03 10 289 ± 1.8 17 1.69E−03 7 H4H14757P 254 ± 0.7 1131.29E−03 9 267 ± 1   109 2.60E−03 4 H4H14758P 308 ± 1.2 126 2.26E−03 5314 ± 0.5 120 3.39E−03 3 H4H14760P2 311 ± 1.4 94 1.87E−03 6 317 ± 2.1 853.90E−03 3

TABLE 4-11 Binding dissociation rate constant of anti-IL-36R monoclonalantibodies binding to mfIL-36R-MMH in two running buffers performed at25° C. PBS-T, pH 7.4 Running Buffer PBS-T, pH 6.0 Running Buffer 100 nM100 nM mfIL- mfIL- mAb 36R- mAb 36R- Capture MMH Capture MMH Level Boundk_(d) t½ Level Bound k_(d) t½ mAb PID (RU) (RU) (1/s) (min) (RU) (RU)(1/s) (min) H4H14699P2 258 ± 1.5 55 2.04E−03 6 276 ± 0.7 48 3.22E−03 4H4H14700P2 331 ± 1.8 91 8.21E−04 14 350 ± 1.7 80 1.71E−03 7 H4H14706P2295 ± 1.7 80 6.46E−04 18 312 ± 1.5 71 9.65E−04 12 H4H14708P2 270 ± 2  57 7.52E−04 15 281 ± 1.2 47 1.50E−03 8 H4H14709P 282 ± 1.3 57 5.19E−0422 301 ± 0.7 48 1.12E−03 10 H4H14728P 264 ± 2   74 5.44E−04 21 269 ± 1.268 6.16E−04 19 H4H14731P 279 ± 2.2 36 1.37E−03 8 279 ± 1.9 23 1.52E−03 8H4H14732P2 245 ± 0.9 14 7.87E−04 15 253 ± 0.9 12 1.15E−03 10 H4H14734P2229 ± 2.2 9 5.05E−04 23 238 ± 1.2 8 6.31E−04 18 H4H14757P 224 ± 1.8 1NB* NB* 235 ± 0.9 1 NB* NB* H4H14758P 243 ± 0.5 0 NB* NB* 254 ± 1   1NB* NB* H4H14760P2 257 ± 1.9 1 NB* NB* 266 ± 1.2 1 NB* NB* *NB indicatesthat under the current experimental conditions, no binding ofmfIL-36R-MMH to anti-hFc captured anti-IL-36R mAb was observed.

TABLE 4-12 Binding dissociation rate constant of anti-IL-36R monoclonalantibodies binding to mfIL-36R-MMH in two running buffers performed at37° C. PBS-T, pH 7.4 Running Buffer PBS-T, pH 6.0 Running Buffer 100 nM100 nM mfIL- mfIL- mAb 36R- mAb 36R- Capture MMH Capture MMH Level Boundkd t½ Level Bound kd t½ mAb PID (RU) (RU) (1/s) (min) (RU) (RU) (1/s)(min) H4H14699P2 310 ± 4.9 58 4.59E−03 3 308 ± 1.9 50 6.22E−03 2H4H14700P2 422 ± 1.1 108 1.80E−03 6 434 ± 1.8 97 3.33E−03 3 H4H14706P2366 ± 1.3 95 1.38E−03 8 375 ± 1.6 85 2.64E−03 4 H4H14708P2 302 ± 3.6 661.69E−03 7 314 ± 2.2 58 3.10E−03 4 H4H14709P 370 ± 2.3 92 9.87E−04 12379 ± 1   80 2.94E−03 4 H4H14728P 305 ± 2.5 98 1.39E−03 8 301 ± 1.2 911.91E−03 6 H4H14731P 266 ± 4   43 3.61E−03 3 279 ± 1.8 40 5.38E−03 2H4H14732P2 302 ± 1.4 18 9.37E−04 12 309 ± 1.7 16 1.57E−03 7 H4H14734P2283 ± 0.8 9 7.87E−04 15 287 ± 1.9 7 1.19E−03 10 H4H14757P 255 ± 0.5 0NB* NB* 267 ± 1.9 −1 NB* NB* H4H14758P 306 ± 1   0 NB* NB* 314 ± 2.5 1NB* NB* H4H14760P2 309 ± 1.6 1 NB* NB* 315 ± 1.3 1 NB* NB* *NB indicatesthat under the current experimental conditions, no binding ofmfIL-36R-MMH to anti-hFc captured anti-IL-36R mAb was observed

Example 5: In Vivo Evaluation of Anti-IL36R in IMQ-Induced SkinInflammation and Chronic Colitis Mouse Models

The anti-human IL-36R monoclonal antibodies of the present inventionwere tested in vivo in acute and chronic Imiquimod (IMQ)-induced skininflammation, and chronic dextran sodium sulfate (DSS)-induced colitisin humanized IL-36R/hIL-36α, β, γ mice. Cytokine detection was performedin skin and colon homogenates using a proinflammatory panel 1 (mouse)multiplex immunoassay kit. Detection of Lipocalin 2 (Lcn2) in fecalhomogenates was performed using a mouse Duoset Lipocalin-2/NGAL EUSAkit. Measurement of myeloperoxidase (MPO) activity in the colonhomogenates was done using a mouse MPO ELISA kit.

The anti-IL36R antibodies, H4H14706P2 and H4H14708P2, were used alongwith a human, isotype matched control IgG4 antibody.

To examine the role of IL-36R in skin and intestinal inflammation and totest the efficacy of hIL-36R antagonism in vivo, anti-human IL-36Rmonoclonal antibodies of the present invention were tested in the murinemodels of Imiquimod (IMQ)-induced skin inflammation and DSS-inducedchronic colitis. In both models, Velocigene generated homozygous miceexpressing human IL-36R and human IL-36α, β, γ and endogenous mouseIL-36Ra were utilized (resulting mice are referred to as DITRA-likemice, due to decreased affinity of mouse IL-36Ra for human IL-36R whichresembles the mutation observed in DITRA (Deficiency of InterleukinThirty-six Receptor Antagonist) patients (Marrakchi et al.,Interleukin-36-receptor antagonist deficiency and generalized pustularpsoriasis, N Engl J Med 365:620-628 (2011)).

A mouse humanized strain with the genotype II1rl2^(hu/hu) II1f6^(hu/hu)II1f8^(hu/hu) II1f9^(hu/hu) was generated. In this mouse strain, humanIL1F6, IL1F8, and IL1F9 replaced the endogenous mouse IL1F6, IL1F8, andIL1F9 (also called IL36α, β and γ respectively); and a chimeric IL1RL2replaced the endogenous mouse IL1RL2. The chimeric IL1RL2 had a humanIL1RL2 extracellular domain and a mouse intracellular domain. Thisresulted in a chimeric receptor that maintained the intracellularsignaling specificity of mice, while rendering the extracellular domainhuman and, thus, able to bind to the human ligands IL1F6, IL1F8, andIL1F9.

Acute and Chronic IMQ-Induced Skin Inflammation Induction and AntibodyTreatment in DITRA-Like Mice.

To induce skin inflammation, 8-10 weeks old humanized DITRA-like femalemice had their back hair shaved using mouse hair trimmer (Oster,MiniMax, Cat #78049-100) and skin depilated with 0.5 g Veet hair removalgel three days prior to IMQ cream application. A daily topical dose of62.5 mg of commercially available IMQ cream (5%) (Aldara, GM Health CareLimited, NDC 99207-206-12) or Vaseline (CVS Pharmacy) was applied on theshaved back skin of the mice for four consecutive days for acute andnine days for chronic disease induction. A daily topical dose of 62.5 mgof Aldara translated into a daily dose of 3.125 mg of an activecompound. In acute IMQ-induced skin inflammation, anti-human IL-36Rantibodies, H4H14706P2 and H4H14708P2, were administered subcutaneouslyinto back skin at 10 mg/kg and 1 mg/kg on three days before (−3d) andone day after (d1) starting the IMQ application. Control group receivedPBS and 10 mg/kg of hIgG4 Isotype control injections. In chronicIMQ-induced skin inflammation, anti-human IL-36R antibodies, H4H14706P2and H4H14708P2, were administered subcutaneously into back skin at 10mg/kg therapeutically on d4 and d8. Two or three days after the start ofIMQ application, the back skin of the mice started to display signs oferythema, scaling and thickening. The severity of inflammation wasmeasured on a daily basis using an adapted version of the clinicalPsoriasis Area and Severity Index. Erythema, scaling and thickening werescored independently on a scale from 0-4: 0, none; 1, slight; 2,moderate; 3, marked; and 4, very marked (van der Fits et al.,Imiquimod-induced psoriasis-like skin inflammation in mice is mediatedvia the IL-23/IL-17 axis. J Immunol 2009, 182:5836-5845). On d4 of acuteand d11 of chronic IMQ-induced skin inflammation, skin thickness wasmeasured using a caliper (Kaefer).

Histopathology.

Skin tissues of 6 mm diameter from murine back were fixed in 10%buffered formalin, and 4-5 μm paraffin embedded sections were stainedwith hematoxylin and eosin. Skin sections were evaluated blindly for thepresence of parakeratosis, orthokeratosis, Munro's microabscess,acanthosis, epidermal ulceration, inflammation in the dermis andhypodermis, blood vessel congestion in the dermis and hypodermis,follicular hyperkeratosis and epithelial hyperplasia. A 0-4 scoringscale was used: 0-within normal limits, 1-minimal, 2-mild, 3-moderateand 4-severe. A total pathology score was calculated for each mouse byadding the individual histopathological feature scores. Data analysiswas performed using GraphPad Prism™ software. Danilenko, Review paper:preclinical models of psoriasis, Vet Pathol. 2008 July; 45(4):563-75;Lowes et al., Pathogenesis and therapy of psoriasis, Nature. 2007 Feb.22; 445(7130):866-73; Mecklenburg et al., Proliferative andnon-proliferative lesions of the rat and mouse integument, J ToxicolPathol. 2013; 26(3 Suppl):27S-57S; Uribe-Herranz et al., IL-1R1signaling facilitates Munro's microabscess formation in psoriasiformimiquimod-induced skin inflammation, J Invest Dermatol. 2013 June;133(6):1541-9; van der Fits et al., Imiquimod-induced psoriasis-likeskin inflammation in mice is mediated via the IL-23/IL-17 axis, JImmunol. 2009 May 1; 182(9):5836-45.

Measurement of Cytokines in Skin Homogenates.

Full thickness skin tissues of 6 mm diameter from murine back were takenand placed in 15 mL tube containing T-per buffer (Thermo Scientific, Cat#378510), 1× Halt Protease Inhibitor Cocktail (Thermo Scientific, Cat#87786) and 5 M EDTA Solution (Thermo Scientific, Cat3 78429). Skintissues were disrupted at 28000 rpm for 1 minute using a Polytron(PT10-35 GT-D, Cat #9158158) and put on ice. Generated skin homogenateswere centrifuged at 1500 rpm for 8 minutes at 4° C. and the supernatantswere collected into 96-well plates. Skin homogenates were subjected toBradford protein assay using protein assay dye (BioRad, Cat #500-0006)to quantify the total protein content. Cytokine concentrations in theskin homogenates were measured using a Proinflammatory Panel 1 (mouse)multiplex immunoassay kit (MesoScale Discovery, Cat # K15048D) accordingto manufacturer's instructions. In brief, 50 μL/well of calibrators andsamples (diluted in Diluent 41) were added to the plates pre-coated withcapture antibodies and incubated at room temperature while shaking at700 rpm for 2 hours. The plates were then washed 3 times with 1×PBScontaining 0.05% (w/v) Tween-20, followed by the addition of 25 μL ofDetection Antibody Solution diluted in Diluent 45. After 2-hoursincubation at room temperature while shaking, the plates were washed 3times, and 150 μL of 2× Read Buffer was added to each well.Electrochemiluminescence was immediately read on a MSD Spector®instrument. Data analysis was performed using GraphPad Prism™ software.Cytokine levels were normalized to total protein content.

Induction of DSS-Induced Model of Chronic Colitis and Antibody Treatmentin DITRA-Like Mice.

To induce chronic DSS-mediated colitis, female DITRA-like mice aged12-20 weeks with an average body weight of more than 23 g were given 3%DSS (Sigma-Aldrich Cat #87786) in drinking water for 7 days followed bydistilled water for 10 days. This cycle was repeated two times untild28. Control group received distilled water for the duration of thestudy. Anti-human IL-36R antibodies, H4H14706P2 and H4H14708P2, wereadministered intraperitoneally at 10 mg/kg and 5 mg/kg bi-weeklystarting on d7. Control group received PBS and 10 mg/kg of hIgG4 Isotypecontrol injections. Mice were weighted and monitored for clinical signsof colitis (e.g., stool consistency and fecal blood) on a daily basis.On d28, mice were euthanized and colon lengths were measured.

Measurement of Lcn-2 in Colon Homogenates.

To monitor intestinal inflammation throughout the study, feces fromindividual DITRA-like mice were collected into 2 mL deep well plates ona weekly basis and stored at −80° C. Upon the completion of the study,feces collected on different days were subjected to homogenization. Inbrief, fecal samples were reconstituted with 1 mL PBS containing 0.1%Tween-20, 1× Halt Protease Inhibitor Cocktail (Thermo Scientific, Cat#87786) and 5 M EDTA Solution (Thermo Scientific, Cat3 78429). Afteradding 2 Tungsten 3 mm Carbide Beads to the wells (Qiagen, Cat #69997),the plates were placed on a shaker at highest speed overnight at 4° C.Homogenous fecal suspensions were centrifuged at 1200 rpm for 10 minutesat 4° C. and the supernatants were collected into 96-well plates. FecalLipocalin-2 (Lcn2) levels were measured using mouse DuosetLipocalin-2/NGAL ELISA kit (R&D Systems, Cat # DY1857) according tomanufacturer's instructions. Data analysis was performed using GraphPadPrism™ software.

Measurement of Myeloperoxidase (MPO) Activity in Colon Homogenates.

Pieces of the distal part of the colon were taken into 2 mLmicrocentrifuge tubes containing 2 Tungsten 3 mm Carbide Beads (Qiagen,Cat #69997) containing T-per buffer (Thermo Scientific, Cat #378510), 1×Halt Protease Inhibitor Cocktail (Thermo Scientific, Cat #87786) and 5 MEDTA Solution (Thermo Scientific, Cat #78429). Colon tissues weredisrupted using Qiagen Tissue Lyser II at a frequency of 27.5 s⁻¹ for 10minutes. Tubes were centrifuged at 1500 rpm for 8 minutes at 4° C. andthe supernatants were collected into 96-well plates. Colon homogenateswere subjected to Bradford protein assay using protein assay dye(BioRad, Cat #500-0006) to quantify the total protein content.Myeloperoxidase (MPO) activity in the colon homogenates was measuredusing mouse MPO ELISA Kit (Hycult Biotech, Cat # HK210-02) according tomanufacturer's instructions. Data analysis was performed using GraphPadPrism™ software. MPO levels were normalized to total protein content

Measurement of Cytokines in Colon Homogenates.

Cytokine concentrations in the colon homogenates were measured using aProinflammatory Panel 1 (mouse) multiplex immunoassay kit (MesoScaleDiscovery, Cat # K15048D) according to manufacturer's instructions. Inbrief, 50 μL/well of calibrators and samples (diluted in Diluent 41)were added to the plates pre-coated with capture antibodies andincubated at room temperature while shaking at 700 rpm for 2 hours. Theplates were then washed 3 times with 1×PBS containing 0.05% (w/v)Tween-20, followed by the addition of 25 μL of Detection AntibodySolution diluted in Diluent 45. After 2-hour incubation at roomtemperature while shaking, the plates were washed 3 times, and 150 μL of2× Read Buffer was added to each well. Electrochemiluminescence wasimmediately read on a MSD Spector® instrument. Data analysis wasperformed using GraphPad Prism™ software. Cytokine levels werenormalized to total protein content

Statistical Analysis.

Statistical significance within the groups was determined by one-wayAnova with Tukey's multiple comparison post-test (^(#), *p<0.01;^(##),**p<0.001; ^(###),***p<0.001; ^(####),****p<0.0001).

Results Summary and Conclusions

Anti-Human IL-36R Monoclonal Antibodies Inhibit Acute Skin Inflammationin DITRA-Like Mice at Prophylactic Dosing.

To examine the role of IL-36R in skin inflammation, two anti-humanIL-36R monoclonal antibodies, H4H14706P2 and H4H14708P2, were tested inIMQ-induced model of psoriasiform dermititis that closely resembleshuman psoriasis lesions in terms of the phenotypic and histologicalcharacteristics (van der Fits et al., Imiquimod-induced psoriasis-likeskin inflammation in mice is mediated via the IL-23/IL-17 axis, JImmunol 2009, 182:5836-5845; Swindell et al., Genome-wide expressionprofiling of five mouse models identifies similarities and differenceswith human psoriasis, PLoS One 2011, 6: e18266; Okayasu et al., A novelmodel in the induction of reliable experimental and chronic ulcerativecolitis in mice, Gastroenterology 1990, 98:694-702). IMQ was applieddaily to the shaved back skin of DITRA-like mice for four consecutivedays. H4H14706P2 and H4H14708P2 antibodies were administered at 10 mg/kgand 1 mg/kg on −3d and d1. Control groups received PBS and hIgG4 Isotypecontrol injections at 10 mg/kg. On d4, skin thickness was measured andtissue harvested for subsequent histopathological evaluation and proteinisolation. Both H4H14706P2 and H4H14708P2 antibodies significantlydecreased IMQ-induced skin thickness in a dose dependent manner comparedto Isotype control (Table 5-1). Histopathological evaluation of the skinlesions revealed a significant reduction in total pathology scoreincluding parakeratosis and Munro's microabscess with anti-human IL-36Rantibody treatment (Table 5-2).

TABLE 5-1 Anti-human IL-36R antibodies reduced skin thickness in acuteIMQ-induced skin inflammation. Thickness is presented in μm. Statisticalsignificance within the groups was determined by one-way Anova withTukey's multiple comparison post-test and standard error of mean (SEM±)calculated: ^(#)significantly different from Vaseline-treated group;*significantly different from PBS- and Isotype-treated groups. N =9/group. IMQ Vaseline H4H14706P2 H4H14708P2 hIgG4 Isotype PBS PBS 1mg/kg 10 mg/kg 1 mg/kg 10 mg/kg 10 mg/kg 496.7 ± 8.8 825 ± 30^(####)674.4 ± 56*** 546.7 ± 30.3**** 624.4 ± 67*** 586.7 ± 53**** 822 ±29.6^(####) p value: ^(#,)*p < 0.01; ^(##,)**p< 0.001; ^(###,)***p <0.001; ^(####,)****p < 0.0001

TABLE 5-2 Anti-human IL-36R antibodies reduced total pathology score inacute IMQ-induced skin inflammation. Statistical significance within thegroups was determined by one-way Anova with Tukey's multiple comparisonpost-test and standard error of mean (SEM±) calculated:^(#)significantly different from Vaseline-treated group; *significantlydifferent from PBS- and Isotype-treated groups. N = 9/group. IMQVaseline H4H14706P2 H4H14708P2 hIgG4 Isotype PBS PBS 1 mg/kg 10 mg/kg 1mg/kg 10 mg/kg 10 mg/kg 0 20.3 ± 3^(####) 17.1 ± 2.4 7 ± 3.6**** 14.2 ±2* 8.9 ± 2.4**** 20.6 ± 1.7^(####) p value: ^(#,)*p < 0.01; ^(##,)**p <0.001; ^(###,)***p < 0.001; ^(####,)****p < 0.0001

Additionally, hIL-36R blockade, with H4H14706P2 and H4H14708P2antibodies, resulted in 66-93% reduction in KC-GRO, IL-6, IL-1β and TNFαproduction in skin homogenates (Table 5-3).

TABLE 5-3 hIL-36R antagonism significantly reduced pro-inflammatorycytokines in IMQ- treated skin of DITRA-like mice (acute skininflammation model). Cytokine levels in PBS/Vaseline control groups weresubtracted from all the treatment groups. Statistical significancewithin the groups was determined by one-way Anova with Tukey's multiplecomparison post-test and standard error of mean (SEM±) calculated:*significantly different from PBS- and Isotype-treated groups. N =9/group. Cytokines IMQ (pg per mg of H4H14706P2 H4H14708P2 hIgG4 Isotypetotal protein) PBS 1 mg/kg 10 mg/kg 1 mg/kg 10 mg/kg 10 mg/kg KC-GRO122.5 ± 31.5 41.5 ± 12.4**** 13.7 ± 4.3**** 35.2 ± 16.7**** 32.3 ±23.5**** 80.4 ± 12.9 IL-6 134.8 ± 13   31.9 ± 12.4**** 18.8 ± 8.4****42.6 ± 17.7**** 37.8 ± 26.9**** 143.5 ± 57.5  IL-1□  84.4 ± 15.2 18.5 ±10.1****  4.9 ± 3.7**** 17.5 ± 13.8**** 7.4 ± 5.6**** 68.1 ± 15.1 TNF-□87.8 ± 6.5 23.6 ± 7.4****   8.2 ± 3.7**** 18.9 ± 8****   9.5 ± 4.4****80.3 ± 15.8 p value: ^(#,)*p < 0.01; ^(##,)**p < 0.001; ^(###,)***p <0.001; ^(####,)****p < 0.0001

Anti-Human IL-36R Monoclonal Antibodies Inhibit Chronic SkinInflammation at Therapeutic Dosing.

To further examine the therapeutic efficacy of hIL-36R antagonism invivo, anti-human IL-36R antibodies were tested in chronic IMQ-inducedmodel of skin inflammation. For the duration of two weeks, IMQ wasapplied to the shaved back skin of DITRA-like mice for nine daysseparated by two days without the application. H4H14706P2 and H4H14708P2antibodies were administered subcutaneously at d4 and d8 at 10 mg/kgdose. Control groups received PBS and hIgG4 Isotype control injectionsat 10 mg/kg. On d1 skin thickness was measured and tissue harvested forsubsequent histopathological evaluation and protein isolation. As shownin Tables 5-4 and 5-5, H4H14706P2 and H4H14708P2 antibodies showedsignificant and comparable efficacy in reducing IMQ-induced skinthickness and pathology lesion scores in DITRA-like mice. Therapeuticadministration of H4H14706P2 and H4H14708P2 led to a significantinhibition of IMQ-induced production of pro-inflammatory cytokines inthe skin of DITRA-like mice (Table 5-6).

TABLE 5-4 Therapeutic administration of anti-human IL-36R antibodiesreduced skin thickness in chronic IMQ-induced skin inflammation. Vase-IMQ line hIgG4 PBS PBS H4H14706P2 H4H14708P2 Isotype 505 ± 70 953 ±74^(####) 667 ± 50**** 674 ± 38**** 951 ± 56.7^(####) Thickness ispresented in μm. Statistical significance within the groups wasdetermined by one-way Anova with Tukey's multiple comparison post-testand standard error of mean (SEM±) calculated: ^(#)significantlydifferent from Vaseline-treated group; *significantly different fromPBS- and Isotype-treated groups. N = 9/group. p value: ^(#,)*p < 0.01;^(##,)**p < 0.001; ^(###,)***p < 0.001; ^(####,)****p < 0.0001

TABLE 5-5 Therapeutic administration of anti-human IL-36R antibodiesreduced total pathology score in chronic IMQ-induced skin inflammation.Vase- IMQ line hIgG4 PBS PBS H4H14706P2 H4H14708P2 Isotype 0 17.2 ±2.9^(####) 12.5 ± 2.2* 9.6 ± 1.9*** 18 ± 2.7^(####) Statisticalsignificance within the groups was determined by one-way Anova withTukey's multiple comparison post-test and standard error of mean (SEM±)calculated: ^(#)significantly different from Vaseline-treated group;*significantly different from PBS- and Isotype-treated groups. N =9/group. p value: ^(#,)*p < 0.01; ^(##,)**p < 0.001; ^(###,)***p <0.001; ^(####,)****p < 0.0001

TABLE 5-6 hIL-36R antagonism significantly inhibited pro-inflammatorycytokines in chronic IMQ-induced skin inflammation. Cytokine levels inPBS/Vaseline control groups were subtracted from all the treatmentgroups. IMQ hIgG4 PBS H4H14706P2 H4H14708P2 Isotype KC-GRO 4.5 ± 1.7 0.6± 0.3**** 0.8 ± 0.4**** 4.4 ± 2.9 IL-6 21.1 ± 6.7  5.1 ± 1****  6.8 ±1.4**** 22.9 ± 13.9 IL-1□ 29.4 ± 11.6 1.8 ± 0.9**** 1.9 ± 0.7**** 23.6 ±19.4 TNF-□  12 ± 2.7 2.1 ± 0.8**** 1.8 ± 0.4**** 14.4 ± 9.3  Statisticalsignificance within the groups was determined by one-way Anova withTukey's multiple comparison post-test and standard error of mean (SEM±)calculated: *significantly different from PBS- and Isotype-treatedgroups. N = 9/group. p value: ^(#,)*p < 0.01; ^(##,)**p < 0.001;^(###,)***p < 0.001; ^(####,)****p < 0.0001

Altogether, these data demonstrated prophylactic and therapeuticefficacy of anti-human IL-36R antibodies in ameliorating IMQ-inducedskin inflammation in vivo. H4H14706P2 and H4H14708P2 antibodiesdisplayed comparable ability to significantly reduce both acute andchronic IMQ-induced skin pathology in DITRA-like mice.

Anti-Human IL-36R Monoclonal Antibodies Ameliorate DSS-Induced ChronicColitis in DITRA-Like Mice at Therapeutic Dosing.

To explore the role of IL-36R antagonism in intestinal inflammation, achemical model of intestinal injury was used. This model utilized oraladministration of DSS that damaged the colonic epithelium (Okayasu etal., A novel model in the induction of reliable experimental and chroniculcerative colitis in mice, Gastroenterology 1990, 98:694-702) andtriggered potent inflammatory responses (Rakoff-Nahoum et al.,Recognition of commensal microflora by toll-like receptors is requiredfor intestinal homeostasis. Cell 2004, 118: 229-241) exhibiting the mainfeatures of IBD-in particular ulcerative colitis. DITRA-like mice weresubjected to chronic DSS-induced colitis by administration of 2-3% DSSfor 7 days followed by 10 days of water for two cycles. H4H14706P2 andH4H14708P2 antibodies were administered at 10 mg/kg and 5 mg/kgbi-weekly starting on d7. Control groups received PBS and hIgG4 Isotypecontrol intraperitoneal injections at 10 mg/kg. To monitor intestinalinflammation at different stages of the disease, feces from individualmice were collected on a weekly basis to measure fecal Lipocalin-2(Lcn2) protein, a non-invasive biomarker of inflammation in intestinalinjury (Thorsvik et al., Fecal neutrophil gelatinase-associatedlipocalin as a biomarker for inflammatory bowel disease. J GastroenterolHepatol 2017, 32:128-135). As shown in Table 5-7, PBS- and hIgG4-treatedgroups displayed significant upregulation of fecal Lcn2 levels on d12,19 (not shown) and 28 compared to water alone. On the contrary, twotherapeutic administrations of H4H14706P2 and H4H14708P2 resulted in asignificant reduction in Lcn2 levels in a dose-dependent manner on d12compared to PBS- and Isotype-treated groups. Sustained reduction offecal Lcn2 levels was observed in anti-human IL-36 antibody-treatedgroups at d19 (not shown) and d28 supporting a role for anti-IL-36Rantibodies in reducing intestinal inflammation in DITRA-like mice (Table5-7). H4H14706P2 antibody displayed better ability to reduce Lcn2 levelsand, thus, intestinal inflammation, compared to H4H14708P2 (Table 5-7).

TABLE 5-7 hIL-36R antagonism significantly reduced fecal Lcn2 levels inDITRA-like mice in chronic DSS-induced colitis. Statistical significancewithin the groups was determined by one- way Anova with Tukey's multiplecomparison post-test and standard error of mean (SEM±) calculated:^(#)significantly different from water-treated group; *significantlydifferent from PBS- and Isotype-treated groups. N = 6-8/group. DSS Day12 Water H4H14706P2 H4H14708P2 hIg4 Isotype d0 d28 d0 PBS 10 mg/kg 5mg/kg 10 mg/kg 5 mg/kg 10 mg/kg 0 0 0 1502 ± 525^(####) 332 ± 107****544 ± 153**** 698 ± 272* 791 ± 5.7 1879 ± 138^(####) DSS Day 28 WaterH4H14706P2 H4H14708P2 hIg4 Isotype d0 d28 PBS 10 mg/kg 5 mg/kg 10 mg/kg5 mg/kg 10 mg/kg 0 0 1379 ± 390^(###) 325 ± 134** 373 ± 217* 635 ± 141600 ± 23* 1448 ± 386^(###) p value: ^(#,)*p < 0.01; ^(##,)**p < 0.001;^(###,)***p < 0.001; ^(####,)****p < 0.0001

hIL-36R blockade with H4H14706P2 and H4H14708P2 antibodies led to adecrease in MPO activity (Table 5-8) and 61-95% reduction inpro-inflammatory cytokines (Table 5-9) in the colon DSS-treatedDITRA-like mice.

TABLE 5-8 Therapeutic administration of anti-human IL-36R antibodiesdecreased MPO activity in the colon of DSS-treated DITRA-like mice. MPOlevels are presented as ng per mg of total protein. Statisticalsignificance within the groups was determined by one-way Anova withTukey's multiple comparison post-test and standard error of mean (SEM±)calculated: ^(#)significantly different from water-treated group;*significantly different from PBS- and Isotype- treated groups. N =6-8/group. DSS Water H4H14706P2 H4H14708P2 hIgG4 Isotype PBS PBS 10mg/kg 5 mg/kg 10 mg/kg 5 mg/kg 10 mg/kg 0 69 ± 19^(####) 6.1 ± 2.3***20.5 ± 6.1*** 29.6 ± 7.5** 23.5 ± 12.7** 64.7 ± 5.6^(###) p value:^(#,)*p < 0.01; ^(##,)**p < 0.001; ^(###,)***p < 0.001; ^(####,)****p <0.0001

TABLE 5-9 Therapeutic administration of anti-human IL-36R antibodiesdecreased pro- inflammatory cytokines in the colon of DSS-treatedDITRA-like mice. MPO levels are presented as ng per mg of total protein.Statistical significance within the groups was determined by one-wayAnova with Tukey's multiple comparison post-test and standard error ofmean (SEM±) calculated: ^(#)significantly different from water-treatedgroup; *significantly different from PBS- and Isotype-treated groups. N= 6-8/group. Cytokines (pg per DSS mg of Water H4H14706P2 H4H14708P2hIgG4 Isotype total protein) PBS PBS 10 mg/kg 5 mg/kg 10 mg/kg 5 mg/kg10 mg/kg KC-GRO 0.54 ± 0.3 98 ± 30^(####) 14.7 ± 3*** 22.9 ± 8**   38.2± 26*  26.4 ± 0.7*   110 ± 12^(###) IL-6 0.69 ± 0.2 345 ± 155^(####) 17.1 ± 6**** 93 ± 75**   69 ± 11***  59 ± 3.4***  627 ± 250^(###) IL-1□ 1.2 ± 0.3 128 ± 17^(####)  13.4 ± 6*** 27 ± 11** 42 ± 46*  34 ± 11* 125 ± 22^(###) TNF-□ 0.98 ± 0.3 74 ± 14^(####)    9.2 ± 4.7****   16 ±8.8****  7.7 ± 4****  4.3 ± 2.1****  28 ± 15^(###) p value: ^(#,)*p <0.01; ^(##,)**p < 0.001; ^(###,)***p < 0.001; ^(####,)****p < 0.0001

Consistent with observations of more reduced Lcn2 levels, H4H14706P2antibody displayed superior efficacy in reducing MPO activity andpro-inflammatory cytokines in the colon compared to H4H14708P2.

Example 6. Epitope Mapping of H4H14706P2, H4H14708P2, and H4H14731PBinding to IL-36R by Hydrogen Deuterium Exchange

Hydrogen Deuterium exchange epitope mapping with mass spectrometry(HDX-MS) was performed to determine the amino acid residues of IL-36R (arecombinant human IL-36R designated as hIL-36R.mmH and having the aminoacid sequence as set forth in SEQ ID NO: 227) interacting withH4H14706P2, H4H14708P2, and H4H14731P (anti-hIL-36R monoclonalantibodies). A general description of the H/D exchange method is setforth in e.g., Ehring (1999) Analytical Biochemistry 267(2):252-259: andEngen and Smith (2001) Anal. Chem. 73:256A-265A.

The HDX-MS experiments were performed on an integrated HDX/MS platform,consisting of a Leaptec HDX PAL system for the deuterium labeling andquenching, a Waters Acquity M-Class (Auxiliary solvent manager) for thesample digestion and loading, a Waters Acquity M-Class (μBinary solventmanager) for the analytical gradient, and Thermo Q Exactive HF massspectrometer for peptide mass measurement.

The labeling solution was prepared as PBS buffer in D₂O at pD 7.0 (10 mMphosphate buffer, 140 mM NaCl, and 3 mM KCl, equivalent to pH 7.4 at 25°C.). For deuterium labeling, 11 μL of IL-36R.mmH (REGN2105, 45.6 pM inH4H14706P2 and H4H14708P2 experiments, or 63.3 μM in H4H14731Pexperiment) or IL-36R.mmH premixed with H4H14706P2, H4H14708P2, orH4H14731P in 1:0.7 molar ratio (Ag-Ab complex) was incubated at 20° C.with 44 μL D₂O labeling solution for various time-points in duplicate(e.g., Undeuterated control=0 second; deuterium-labeled for 5 minutesand 10 minutes). The deuteration reaction was quenched by adding 55 μLof pre-chilled quench buffer (0.5 M TCEP-HCl, 8 M urea and 1% formicacid) to each sample for a 5-minute incubation at 20° C. The quenchedsample was then injected into a Waters HDX Manager for onlinepepsin/protease XIII digestion. The digested peptides were separated bya C8 column (1.0 mm×50 mm, NovaBioassays) with a 13-minute gradient from10%-32% B (mobile phase A: 0.5% formic acid in water, mobile phase B:0.1% formic acid in acetonitrile). The eluted peptides were analyzed byQ Exactive HF mass spectrometry in LC-MS/MS or LC-MS mode.

The LC-MS/MS data of undeuterated IL-36R sample were searched against adatabase including IL-36R and its randomized sequence using Byonicsearch engine (Protein Metrics). The search parameters (in ELN) were setas default using non-specific enzymatic digestion and humanglycosylation as common variable modification. The list of identifiedpeptides was then imported into the HDX Workbench software (version 3.3)to calculate the deuterium uptake of each peptide detected by LC-MS fromall deuterated samples. For a given peptide, the centroid mass(intensity-weighted average mass) at each time point was used tocalculate the deuterium uptake (D) and percentage of deuterium uptake (%D).

${{Deuterium}\mspace{14mu} {Uptake}\mspace{14mu} \left( {D\text{-}{uptake}} \right)} = \begin{matrix}{{{Average}\mspace{14mu} {Mass}\mspace{14mu} ({deuterated})} - {{Average}\mspace{14mu} {Mass}}} \\({undeuterated})\end{matrix}$${{Percentage}\mspace{14mu} {of}\mspace{14mu} {deuterium}\mspace{14mu} {uptake}\mspace{14mu} \left( {\% \mspace{11mu} D} \right)} = \frac{D\text{-}{update}\mspace{14mu} {for}\mspace{14mu} {peptide}\mspace{14mu} {at}\mspace{14mu} {each}\mspace{14mu} {time}\mspace{14mu} {point} \times 100\%}{{Maximum}\mspace{14mu} D\text{-}{uptake}\mspace{14mu} {of}\mspace{14mu} {the}\mspace{14mu} {peptide}\mspace{14mu} \left( {{defined}\mspace{14mu} {in}\mspace{14mu} {ELN}} \right)}$

A total of 163 peptides from REGN2105 (hIL-36R.mmH) were identified fromboth hIL-36R.mmH alone and hIL-36R.mmH in complex with H4H14706P2samples, representing 81.5% sequence coverage of hIL-36R. Any peptidewhich exhibited a differential percent D-uptake value above 5% wasdefined as significantly protected. Peptides corresponding to aminoacids 113-122 (YKQILHLGKD) (SEQ ID NO: 229) (amino acids 113-122 of SEQID NO: 227) on REGN2105 were significantly protected by H4H14706P2.

A total of 148 peptides from REGN2105 (hIL-36R.mmH) were identified fromboth hIL-36R.mmH alone and hIL-36R.mmH in complex with H4H14708P2samples, representing 80.1% sequence coverage of hIL-36R. Any peptidewhich exhibited a differential percent D-uptake value above 5% wasdefined as significantly protected. Peptides corresponding to aminoacids 113-122 (YKQILHLGKD) (SEQ ID NO: 229) (amino acids 113-122 of SEQID NO: 227) on REGN2105 were significantly protected by H4H14708P2.

A total of 237 peptides from REGN2105 (hIL-36R.mmH) were identified fromboth hIL-36R.mmH alone and hIL-36R.mmH in complex with H4H14731Psamples, representing 88.2% sequence coverage of hIL-36R. Any peptidewhich exhibited a differential percent D-uptake value above 5% wasdefined as significantly protected. Peptides corresponding to aminoacids 264-277 (GVETHVSFREHNLY) (SEQ ID NO: 230) (amino acids 264-277 ofSEQ ID NO: 227) on REGN2105 were significantly protected by H4H14731P.

TABLE 6-1 IL-36R.mmH peptides with significant protection upon bindingto H4H14706P2 5 min 10 min REGN2105 + REGN2105 + REGN2105 H4H14706P2REGN2105 H4H14706P2 IL-36R Charge Centroid Centroid Centroid Residues(+) MH⁺ MH⁺ ΔD Centroid MH⁺ MH⁺ ΔD Δ% D 113-119 2 918.84 918.60 −0.24918.97 918.71 −0.26 −6.2 113-122 1 1218.76 1218.29 −0.47 1218.91 1218.45−0.46 −7.2 113-122 2 1219.97 1219.52 −0.45 1220.10 1219.66 −0.44 −6.9116-119 1 497.13 497.01 −0.12 497.19 497.03 −0.17 −8.9 116-122 1 798.29797.98 −0.31 798.35 797.99 −0.35 −8.3

TABLE 6-2 IL-36R.mmH peptides with significant protection upon bindingto H4H14708P2 5 min 10 min REGN2105 + REGN2105 + REGN2105 H4H14708P2REGN2105 H4H14708P2 IL-36R Charge Centroid Centroid Centroid CentroidResidues (+) MH⁺ MH⁺ ΔD MH⁺ MH⁺ ΔD Δ% D 113-119 2 918.84 918.59 −0.25918.97 918.69 −0.28 −6.6 113-122 1 1218.73 1218.13 −0.61 1218.90 1218.31−0.58 −9.3 113-122 2 1219.97 1219.51 −0.46 1220.10 1219.58 −0.51 −7.6116-119 1 497.13 497.01 −0.12 497.19 497.03 −0.17 −9.0 116-122 1 798.29797.90 −0.39 798.35 797.93 −0.42 −10.1

TABLE 6-3 IL-36R.mmH peptides with significant protection upon bindingto H4H14731P 5 min 10 min REGN2105 + REGN2105 + REGN2105 H4H14731PREGN2105 H4H14731P IL-36R Charge Centroid Centroid Centroid CentroidResidues (+) MH⁺ MH⁺ ΔD MH⁺ MH⁺ ΔD Δ% D 264-271 2 880.18 879.93 −0.24880.24 879.91 −0.32 −5.9 267-271 1 592.76 592.58 −0.18 592.78 592.56−0.23 −8.5 268-271 1 491.44 491.23 −0.21 491.47 491.22 −0.25 −14.4268-276 3 1144.59 1144.25 −0.34 1144.60 1144.21 −0.39 −6.6 268-277 31307.97 1307.56 −0.41 1308.01 1307.48 −0.53 −7.3 271-276 2 818.86 818.67−0.19 818.86 818.63 −0.23 −6.5

Amino acid sequence of recombinant human IL-36R (IL1RL2: interleukin 1receptor-like 2; REGN2105) (hIL36R.mmH): Monomeric human IL-36R (aminoacids D20-Y337, Accession # Q9HB29), with a C-terminalmyc-myc-hexahistidine (mmH) tag (underlined):

(SEQ ID NO: 227) DGCKDIFMKNEILSASQPFAFNCTFPPITSGEVSVTWYKNSSKIPVSKIIQSRIHQDETWILFLPMEWGDSGVYQCVIKGRDSCHRIHVNLTVFEKHWCDTSIGGLPNLSDEYKQILHLGKDDSLTCHLHFPKSCVLGPIKWYKDCNEIKGERFTVLETRLLVSNVSAEDRGNYACQAILTHSGKQYEVLNGITVSITERAGYGGSVPKIIYPKNHSIEVQLGTTLIVDCNVTDTKDNTNLRCWRVNNTLVDDYYDESKRIREGVETHVSFREHNLYTVNITFLEVKMEDYGLPFMCHAGVSTAYIILQLPAPDFRAYEQKLISEEDLGGEQKLISEEDLHHHHHH

Example 7: In Vivo Evaluation of Anti-IL36R in IMQ-Induced andOxazolone-Induced Skin Inflammation and Chronic Colitis Mouse Models

The anti-human IL-36R monoclonal antibodies of the present inventionwere tested in primary human cell assays in vitro; and compared withother anti-human IL-36R monoclonal antibodies in in vivo Imiquimod(IMQ)-induced skin inflammation assays in humanized IL-36R/hIL-36α, β, γmice. Also, the anti-human IL-36R monoclonal antibodies of the presentinvention were tested in vivo in an oxazolone-induced model of colitisin humanized IL-36R/hIL-36α, β, γ mice.

IL-8 was detected in culture supernatants using DuoSet ELISA kit forHuman CXCL8/IL-8 (R&D Systems) and cytokine was detected in skin andcolon homogenates using Proinflammatory Panel 1 (mouse and human)Multiplex Immunoassay kit (MSD). Monoclonal antibodies tested wereH4H14706P2, H4H14708P2, APE6155 (IgG4) and a human IgG4 isotype control(REGN1002).

The APE6155 heavy chain (comprising an IgG4 constant domain) comprisesthe amino acid sequence:

(SEQ ID NO: 239) QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYWMNWVRQAPRQGLEWMGMFHPTGDVTRLNQKFKDRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARTTSMIIGGFAYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSVSVMHEALHNHYTQKSLSLSLGK

The APE6155 light chain (comprising a Kappa constant domain) comprisesthe amino acid sequence:

(SEQ ID NO: 240) DIVMTQTPLSLSVTPGQPASISCRSSKSLLHRNAITYFYWYLHKPGQPPQLLIYQMSNLASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCAQNLELPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

See WO2016/168542.

Testing Anti-Human IL-36R Antibodies In Vitro in Primary Human CellsAssays In Vitro.

Normal Human Epidermal Keratinocytes (NHLF; Lonza, Cat #00192627, lot#254498) and Intestinal MyoFibroblast (InMyoFib; Lonza, Cat # CC-2902,lot #0000254498) were cultured in vitro for 4-5 passages in KGM-Gold™supplemented with BulletKit™ (Lonza, Cat # CC-00192060, lot #0000484385)and SmGmm-2 supplemented with BulletKit™ (Lonza, Cat # CC-3182, lot#00004736694), respectively. Human CD14+ monocytes were isolated fromperipheral blood of 3 different donors using EasySep Human MonocyteIsolation Kit (StemCell, Cat #19359) per manufacturer's instructions. Aday before the assay, primary human cells were plated in correspondingMedia at 10000 per well in 96-well flat bottom plate and incubatedovernight at 37° C. Cells were stimulated in the presence of constantconcentration (10 nM) or serially diluted (starting from 1500 nM)rhIL-36α/IL-1F6 [aa6-158] (R&D Systems, Cat #6995-IL-010/CF, lot #DAFZ0313051), rhIL-363/IL-1F8 [aa5-157] (R&D Systems, Cat#6834-IL-010/CF, lot # DAKU0514062 and rhIL-36γ/IL-1F9 [aa18-169] (R&DSystems, Cat #6835-IL-010/CF, lot # DAPK0215011) alone or incombination. Serial dilutions starting from 2400 nM of anti-human IL-36Rantibodies were added to the wells. Plates were incubated for 24 hoursat 37° C. and supernatants were collected to measure IL-8 using DuoSetELISA Development System for Human CXCL8/IL-8 (R&D Systems, Cat #DY208-05, lot #325963). To obtain EC₅₀ and IC₅₀ values, the results wereanalyzed using nonlinear regression (4-parameter logistics) in GraphPadPrism™ software.

Testing and Comparing Anti-Human IL-36R Antibodies in IMQ-Induced SkinInflammation.

To induce skin inflammation, 8-10 weeks old humanized DITRA-like femalemice had their back hair shaved using mouse hair trimmer (Oster,MiniMax, Cat #78049-100) and skin depilated with 0.5 g Veet hair removalgel three days prior to IMQ cream application. A daily topical dose of62.5 mg of commercially available IMQ cream (5%) (Aldara, GM Health CareLimited, NDC 99207-206-12, lot # QJ044A) or Vaseline (CVS Pharmacy, NDC59779-902-88) was applied on the shaved back skin of the mice for fourconsecutive days. A daily topical dose of 62.5 mg of Aldara translatedinto a daily dose of 3.125 mg of an active compound. Anti-human IL-36Rantibodies—H4H14706P2, H4H14708P2 and APE6155 (IgG4), were administeredsubcutaneously into back skin at 10 mg/kg on −3d and d1. Control groupreceived PBS and 10 mg/kg of hIgG4 Isotype control (REGN1002)injections. Two or three days after the start of IMQ application, theback skin of the mice started to display signs of erythema, scaling andthickening. The severity of inflammation was measured on a daily basisusing an adapted version of the clinical Psoriasis Area and SeverityIndex. Erythema, scaling and thickening were scored independently on ascale from 0-4: 0, none; 1, slight; 2, moderate; 3, marked; and 4, verymarked (van der Fits et al. Imiquimod-induced psoriasis-like skininflammation in mice is mediated via the IL-23/IL-17 axis. J Immunol2009, 182:5836-5845). Skin thickness was measured using caliper on d5(Kaefer).

Histopathology.

Skin tissues of 6 mm diameter from murine back were fixed in 10%buffered formalin, and 4-5 μm paraffin embedded sections were stainedwith hematoxylin and eosin. Skin sections were evaluated blindly for thepresence of parakeratosis, orthokeratosis, Munro's microabscess,acanthosis, epidermal ulceration, inflammation in the dermis andhypodermis, blood vessel congestion in the dermis and hypodermis,follicular hyperkeratosis and epithelial hyperplasia. A 0-4 scoringscale was used: 0-within normal limits, 1-minimal, 2-mild, 3-moderateand 4-severe. A total pathology score was calculated for each mouse byadding the individual histopathological feature scores. Data analysiswas performed using GraphPad Prism™ software.

Measurement of Cytokines in Skin Homogenates.

Full thickness skin tissues of 6 mm diameter from murine back were takenand placed in 15 mL tube containing T-per buffer (Thermo Scientific, Cat#378510, lot # RF236217), 1× Halt Protease Inhibitor Cocktail (ThermoScientific, Cat #87786, lot # QG221763) and 5 M EDTA Solution (ThermoScientific, Cat3 78429). Skin tissues were disrupted at 28000 rpm for 1minute using a Polytron (PT10-35 GT-D, Cat #9158158) and put on ice.Generated skin homogenates were centrifuged at 1500 rpm for 8 minutes at4° C. and the supernatants were collected into 96-well plates. Skinhomogenates were subjected to Bradford protein assay using protein assaydye (BioRad, Cat #500-0006, lot #210008149) to quantify the totalprotein content. Cytokine concentrations in the skin homogenates weremeasured using a Proinflammatory Panel 1 (mouse) multiplex immunoassaykit (MesoScale Discovery, Cat # K15048D) according to manufacturer'sinstructions. In brief, 50 μL/well of calibrators and samples (dilutedin Diluent 41) were added to the plates pre-coated with captureantibodies and incubated at room temperature while shaking at 700 rpmfor 2 hours. The plates were then washed 3 times with 1×PBS containing0.05% (w/v) Tween-20, followed by the addition of 25 μL of DetectionAntibody Solution diluted in Diluent 45. After 2-hours incubation atroom temperature while shaking, the plates were washed 3 times, and 150μL of 2× Read Buffer was added to each well. Electrochemiluminescencewas immediately read on a MSD Spector® instrument. Data analysis wasperformed using GraphPad Prism™ software. Cytokine levels werenormalized to total protein content.

Testing Anti-Human IL-36R Monoclonal Antibodies in Oxazolone-InducedIntestinal Inflammation—Induction of Oxazolone-Induced Model of ChronicColitis and Antibody Treatment in DITRA-Like Mice.

Oxazolone colitis was induced as previously described (Heller et al.,Oxazolone colitis, a Th2 colitis model resembling ulcerative colitis, ismediated by IL-13-producing NK-T cells. Immunity 2002, 17: 629-638).Briefly, in order to pre-sensitize DITRA-like mice, a 2×2 cm² field ofthe abdominal skin was shaved, and 100 μl of a 3% solution oxazolone((4-ethoxymethylene-2-phenyl-2-oxazoline-5-one; Sigma Aldrich) dilutedin 100% ethanol was applied. On days 5 and 7 after pre-sensitization,mice were challenged intrarectally with 50 μl of 1.5% oxazolone dilutedin 50% ethanol under general anesthesia. Control mice werepre-sensitized with 100% ethanol and received intrarectal injection of50% ethanol. Anti-human IL-36R antibodies—H4H14706P2 and H4H14708P2,were administered intraperitoneally at 10 mg/kg on d2, 5 and 7. Controlgroup received PBS and 10 mg/kg of hIgG4 Isotype control (REGN1002)injections. Mice were weighted and monitored for clinical signs ofcolitis (e.g., stool consistency and fecal blood) on a daily basis. Ond8, mice were euthanized and colons were collected.

Measurement of Cytokines in Colon Homogenates.

Pieces of distal part of the colon were taken into 2 mL microcentrifugetubes containing 2 Tungsten 3 mm Carbide Beads (Qiagen) containing T-perbuffer (Thermo Scientific), 1× Halt Protease Inhibitor Cocktail (ThermoScientific) and 5M EDTA Solution (Thermo Scientific). Colon tissues weredisrupted using Qiagen Tissue Lyser II at frequency of 27.5 s⁻¹ for 10minutes. Tubes were centrifuged at 1500 rpm for 8 minutes at 4° C. andthe supernatants were collected into 96-well plates. All tissuehomogenates were subjected to Bradford protein assay using protein assaydye (BioRad) to quantify the total protein content.

Cytokine concentrations in the colon homogenates were measured using aProinflammatory Panel 1 (mouse) multiplex immunoassay kit (MesoScaleDiscovery Cat # K15048D) according to manufacturer's instructions. Inbrief, 50 μL/well of calibrators and samples (diluted in Diluent 41)were added to the plates pre-coated with capture antibodies andincubated at room temperature while shaking at 700 rpm for 2 hours. Theplates were then washed 3 times with 1×PBS containing 0.05% (w/v)Tween-20, followed by the addition of 25 μL of Detection AntibodySolution diluted in Diluent 45. After 2-hour incubation at roomtemperature while shaking, the plates were washed 3 times, and 150 μL of2× Read Buffer was added to each well. Electrochemiluminescence wasimmediately read on MSD Spector® instrument. Data analysis was performedusing GraphPad Prism™ software. Cytokine levels were normalized to totalprotein content.

Statistical Analysis.

Statistical significance within the groups was determined by one-wayAnova with Tukey's multiple comparison post-test (*p<0.05, *p<0.005,***p<0.0005, ****p<0.00001).

Results Summary and Conclusions

Anti-Human IL-36R Monoclonal Antibodies Potently Inhibit Human IL-36RSignaling in Primary Human Cells In Vitro.

Human Epidermal Keratinocytes (NHEK), Human Intestinal Myofibroblasts(InMyoFib) and Peripheral Blood (PB)-derived CD14⁺ Monocytes werestimulated in vitro with 10 nM of IL-36α, β and γ. Serially dilutedanti-human IL-36R monoclonal antibodies (H4H14706P2 and H4H14708P2) wereadded to the cultures, supernatants were collected 24 hourspost-incubation and human IL-8 production in response to IL-36stimulation was measured. The anti-human IL-36R monoclonal antibodiespotently inhibit all three IL-36 cytokines in Human EpidermalKeratinocytes, Human Intestinal Myofibroblasts and Peripheral Blood(PB)-derived CD14⁺ Monocytes in vitro with IC₅₀ 1-6 nM (Table 7-1).

TABLE 7-1 Anti-human IL-36R antibodies, H4H14706P2 and H4H14708P2,inhibited human IL-36α, β and γ in human primary cells in vitro. NormalEpidermal Intestinal MyoFibroblasts PB-derived Cells Keratinocytes(NHEK) (InMyoFib) CD14⁺ Monocytes hIL-36 Ligands α β γ α β γ α β γEC50[M] 1.43E−09 1.18E−09 4.00E−09 1.41E−09 1.18E−09 1.46E−09 2.75E−092.63E−09 2.75E−09 Constant for Inhibition 10 nM 10 nM 10 nM 10 nM 10 nM10 nM 10 nM 10 nM 10 nM Protein/hIL-36R ab IC50 [M] IC50 [M] IC50 [M]IC50 [M] IC50 [M] IC50 [M] IC50 [M] IC50 [M] IC50 [M] H4H14706P24.42E−09 3.62E−09 2.11E−09 4.89E−09 3.62E−09 4.59E−09 1.15E−09 1.74E−091.58E−09 H4H14708P2 5.06E−09 5.79E−09 3.63E−09 5.72E−09  5.3E−096.40E−09 2.38E−09 2.24E−09 1.95E−09 hIgG Ctr (REGN1002) None None NoneNone None None None None None

Anti-Human IL-36R Monoclonal Antibodies H4H14706P2 and H4H14708P2 areMore Potent than the APE6155 Antibody in Inhibiting IMQ-Induced SkinInflammation in DITRA-Like Mice.

H4H14706P2 and H4H14708P2 and APE6155 anti-human IL-36R monoclonalantibodies were tested head-to-head in IMQ-induced model of psoriasiformdermatitis. IMQ was applied daily to the shaved back skin of DITRA-likemice for four consecutive days. H4H14706P2 and H4H14708P2 and APE6155antibodies were administered at 10 mg/kg on −3d and d1. Control groupsreceived PBS and hIgG4 Isotype control injections at 10 mg/kg. On d5,skin thickness was measured and tissue harvested for subsequenthistopathological evaluation and protein isolation. Both H4H14706P2 andH4H14708P2 antibodies displayed greater potency in significantlydecreasing IMQ-induced skin thickness compared to APE6155 (Table 7-2).Histopathological evaluation of the skin lesions revealed a greaterreduction in total pathology score including parakeratosis and Munro'smicroabscess with anti-human IL-36R antibodies treatments (Table 7-3).

TABLE 7-2 Anti-human IL-36R antibodies, H4H14706P2 and H4H14708P2, aremore potent than APE6155 anti-human IL-36R antibody in reducing skinthickness in IMQ-induced skin inflammation.^(~) Vase- IMQ line hIgG 4PBS PBS H4H14706P2 H4H14708P2 APE6155 Isotype 607 ± 18 748 ± 45 586 ±34** 585 ± 24** 689 ± 81 740 ± 42.5 ^(~)Thickness is presented in μm.Statistical significance within the groups was determined by one-wayAnova with Tukey's multiple comparison post-test and standard error ofmean (SEM±) calculated: *significantly different from PBS- andIsotype-treated groups. n = 9/group.

TABLE 7-3 Anti-human IL-36R antibodies, H4H14706P2 and H4H14708P2,displayed greater potency than APE6155 in reducing the total pathologyscore in IMQ-induced skin inflammation.^($) Vase- IMQ line hIgG 4 PBSPBS H4H14706P2 H4H14708P2 APE6155 Isotype 2 ± 0.6 17 ± 2.5 11 ± 1.4***11.6 ± 1.9** 13.4 ± 3 18 ± 1.5 ^($)Statistical significance within thegroups was determined by one-way Anova with Tukey's multiple comparisonpost-test and standard error of mean (SEM±) calculated: *significantlydifferent from PBS- and Isotype-treated groups. n = 9/group.

Additionally, human IL-36R blockade with H4H14706P2 and H4H14708P2antibodies resulted in greater reduction in KC-GRO, 11-6, IL-1β and TNFαproduction in skin homogenates compared to COMP5382 (Table 7-4).

TABLE 7-4 Anti-human IL-36R antibodies, H4H14706P2 and H4H14708P2,displayed greater potency than APE6155 in reducing pro-inflammatorycytokines in the skin.^(∞) Cytokines IMQ (pg per mg of total tissue) PBSH4H14706P2 H4H14708P2 APE6155 hIgG4 Isotype KC-GRO  64 ± 10 19 ± 5.5** 23 ± 7**    40 ± 16  65 ± 22 IL-6 160 ± 47 41 ± 14**** 51 ± 16**** 128 ±59 165 ± 87 L-1β 128 ± 43 8.6 ± 1.9****  10 ± 1.3****    28 ± 17**** 117± 49 THF-α  72 ± 22 11 ± 4.2*   12 ± 2.9*   20.5 ± 9.8  65 ± 22^(∞)Values are presented as “pg per mg of total tissue”. Statisticalsignificance within the groups was determined by one-way Anova withTukey's multiple comparison post-test and standard error of mean (SEM±)calculated: *significantly different from PBS- and Isotype-treatedgroups, n = 9/group.

Anti-Human IL-36R Monoclonal Antibodies Ameliorate Oxazolone-InducedColitis in DITRA-Like Mice.

To further explore biological function of IL-36 in the gut, we testedthe efficacy of IL-36R blockade in oxazolone-induced colitis, anotherpreclinical model of IBD with the histologic resemblance to humanulcerative colitis (Heller et al.). Prophylactic administration ofanti-human IL-36R antibodies, H4H14706P2 and H4H14708P2, significantlyreduced oxazolone-induced disease severity in DITRA-like mice comparedto PBS and isotype control treated groups as reflected in levels ofIL-4, IL-6 and TNF-α in the colon of oxazolone-treated DITRA-like mice(Table 7-5).

TABLE 7-5 Human IL-36R antagonism ameliorates oxazolone-induced colitisin DITRA-like mice in vivo.^(♦) Cytokines Oxazolone (pg per mg of totaltissue) Vehicle PBS H4H14706P2 H4H14708P2 hIgG4 Isotype IL-4  2 ± 0.4509 ± 148  51.5 ± 28**** 111 ± 35**** 296 ± 106 IL-6 34 ± 29 2128 ± 1255198 ± 131** 214 ± 116**  2276 ± 1338 TNF-□ 95 ± 36 822 ± 149 387 ± 126**569 ± 136   859 ± 148 ^(♦)DITRA-like mice were pre-sensitized with 3%solution of oxazolone dissolved in 100% Ethanol and intrarectallyadministered with 1.5% oxazolone and vehicle (50% Ethanol) 5 days later.Mice were intraperitoneally injected with PBS, anti-human IL-36R mAb andhIgG4 Isotype control on days 2, 5 and 7 after pre-sensitization. Levelsof pro-inflammatory cytokines in colon homogenates in oxazolone- andvehicle- treated DITRA-like mice injected with PBS, anti-human IL-36RmAb and hIgG4 Isotype control. Values are presented as “pg per mg oftotal tissue”. Statistical significance within the groups was determinedby one-way Anova with Tukey's multiple comparison post-test and standarderror of mean (SEM±) calculated: *represents significant difference fromPBS-treated group, n = 5/group.

Example 8: Bioassay Using Human HEK293/NFkB-luc/hIL36R Cell Line forSchild Analysis

To characterize the inhibitory properties of the anti-IL36R antibodies,H4H14706P2 and H4H14708P2, a Schild analysis was performed. This methodassesses the nature of antagonism by inhibitors and measures theaffinity of a competitive antagonist when a number of conditions arefulfilled (Colquhoun, Why the Schild method is better than Schildrealized, Trends Pharmacol Sci. 2007 December; 28(12):608-14).

For the bioassay, HEK293/NFκB-luc/hIL-36R cells are seeded onto 96-wellassay plates at 10,000 cells/well in low serum media, 0.1% FBS andOPTIMEM, and incubated at 37° C. and 5% CO₂ overnight. Next day,antibody was added to cells at different, fixed concentrations (9 nM, 3nM, 1 nM, 0.3 nM or 0.1 nM) and pre-incubated with cells for 15 minutesat room temperature. A condition without antibody was also included.IL-36α, IL-36β, or IL-36γ were then serially diluted from 100 nM to 2 pMor 100 nM to 0.1 pM and were added to cells along with sample withoutany ligand. Luciferase activity was detected after 5.5 hrs of incubationin 37° C. and 5% CO₂ with Victor X5 or EnVision™ Multilabel Plate Reader(Perkin Elmer) and the results were analyzed using Gaddum/Schild EC₅₀shift with Prism 7 (GraphPad).

A Schild analysis of H4H14706P2 and H4H14708P2 showed that increasingamount of antibodies caused parallel rightward shift of the IL36 liganddose-response curves and that the inhibitory effect of H4H14706P2 andH4H14708P2 was surmountable by increasing amounts of IL36 ligand,suggesting competitive inhibition of H4H14706P2 and H4H14708P2 (FIGS.3A-3F).

Example 9: Pharmacokinetic (PK) Studies

Female cynomolgus monkeys were assigned to dose groups for PKcharacterization; animals (3 animals/group) received a single SCinjection of 5 or 0.5 mg/kg of H4H14708P2 or APE6155. Blood samples werecollected from all animals at pre-dose and at 4, 24, 48, 72, 120, 168,240, 336, 504, 576, 672, 840, 912, 1008, 1080, 1176, 1248, 1344, 1512and 1680 hours post dose. Concentrations of total H4H14708P2 or APE6155in serum were determined using a non-validated enzyme-linkedimmunosorbent assays (ELISAs). The method was designed to measure totalhuman IgG concentrations in cynomolgus serum. Pharmacokinetic parameterswere estimated using non-compartmental analysis. H4H14708P2 was observedto have about 1.3-fold greater exposure than APE6155 at 5 mg/kg dosageand about 1.2-fold greater exposure than APE6155 at 0.5 mg/kg dosage.See FIG. 4.

All references cited herein are incorporated by reference to the sameextent as if each individual publication, database entry (e.g., Genbanksequences or GeneID entries), patent application, or patent, wasspecifically and individually indicated to be incorporated by reference.This statement of incorporation by reference is intended by Applicants,to relate to each and every individual publication, database entry(e.g., Genbank sequences or GeneID entries), patent application, orpatent even if such citation is not immediately adjacent to a dedicatedstatement of incorporation by reference. The inclusion of dedicatedstatements of incorporation by reference, if any, within thespecification does not in any way weaken this general statement ofincorporation by reference. Citation of the references herein is notintended as an admission that the reference is pertinent prior art, nordoes it constitute any admission as to the contents or date of thesepublications or documents.

1. An isolated antibody or antigen-binding fragment thereof that: bindsto a polypeptide comprising the amino acid sequence set forth in SEQ IDNO: 227 at residues 113-119, 113-122, 116-119, 116-122, 264-271,267-271, 268-271, 268-276, 268-277 and/or 271-276; or, (i) specificallybinds to the same epitope on IL36R as a reference antibody orantigen-binding fragment thereof; or (ii) competes for binding to IL36Rpolypeptide with a reference antibody or antigen-binding fragmentthereof, wherein the reference antibody or antigen-binding fragmentthereof comprises: (a) a heavy chain immunoglobulin or variable regionthereof that comprises CDR-H1, CDR-H2 and CDR-H3 of a heavy chainimmunoglobulin or variable region thereof that comprises the amino acidsequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130,138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or224; or a variant thereof; and/or (b) a light chain immunoglobulin orvariable region thereof that comprises CDR-L1, CDR-L2 and CDR-L3 of alight chain immunoglobulin or variable region thereof that comprises theamino acid sequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106,122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or226; or a variant thereof.
 2. An isolated antibody or antigen-bindingfragment thereof comprising: (i) a heavy chain immunoglobulin orvariable region thereof that comprises CDR-H1, CDR-H2 and CDR-H3 of aheavy chain immunoglobulin or variable region thereof that comprises theamino acid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98,114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212,216, 220 or 224; or a variant thereof; and/or (ii) a light chainimmunoglobulin or variable region thereof that comprises CDR-L1, CDR-L2and CDR-L3 of a light chain immunoglobulin or variable region thereofthat comprises the amino acid sequence set forth in SEQ ID NO: 10, 26,42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202, 206,210, 214, 218, 222 or 226; or a variant thereof.
 3. The antibody orantigen-binding fragment thereof of claim 1 comprising: (a) a heavychain immunoglobulin or variable region thereof comprising an amino acidsequence having at least 90% amino acid sequence identity to the aminoacid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114,130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216,220 or 224; and/or (b) a light chain immunoglobulin or variable regionthereof comprising an amino acid sequence having at least 90% amino acidsequence identity to the amino acid sequence set forth in SEQ ID NO: 10,26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190, 194, 198, 202,206, 210, 214, 218, 222 or
 226. 4. The antibody or antigen-bindingfragment thereof of claim 1 comprising: (a) a heavy chain immunoglobulinor variable region thereof comprising the CDR-H1, CDR-H2 and CDR-H3 of aheavy chain immunoglobulin or variable region thereof comprising anamino acid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98,114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212,216, 220 or 224, and at least 90% amino acid sequence identity to theamino acid sequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98,114, 130, 138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212,216, 220 or 224; and/or (b) an light chain immunoglobulin or variableregion thereof comprising the CDR-L1, CDR-L2 and CDR-L3 of a light chainimmunoglobulin or variable region thereof comprising an amino acidsequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146,162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or 226, andat least 90% amino acid sequence identity to the amino acid sequence setforth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182,186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or
 226. 5. The antibodyor antigen-binding fragment thereof of claim 1 comprising: a heavy chainimmunoglobulin or variable region thereof that comprises: CDR-H1comprising the amino acid sequence set forth in SEQ ID NO: 4, or avariant thereof; CDR-H2 comprising the amino acid sequence set forth inSEQ ID NO: 6, or a variant thereof; and CDR-H3 comprising the amino acidsequence set forth in SEQ ID NO: 8, or a variant thereof and/or CDR-H1comprising the amino acid sequence set forth in SEQ ID NO: 20, or avariant thereof; CDR-H2 comprising the amino acid sequence set forth inSEQ ID NO: 22, or a variant thereof; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 24, or a variant thereof and/orCDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 36, ora variant thereof; CDR-H2 comprising the amino acid sequence set forthin SEQ ID NO: 38, or a variant thereof; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 40, or a variant thereof and/orCDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 52, ora variant thereof; CDR-H2 comprising the amino acid sequence set forthin SEQ ID NO: 54, or a variant thereof; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 56, or a variant thereof and/orCDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 68, ora variant thereof; CDR-H2 comprising the amino acid sequence set forthin SEQ ID NO: 70, or a variant thereof; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 72, or a variant thereof and/orCDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 84, ora variant thereof; CDR-H2 comprising the amino acid sequence set forthin SEQ ID NO: 86, or a variant thereof; and CDR-H3 comprising the aminoacid sequence set forth in SEQ ID NO: 88, or a variant thereof and/orCDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 100,or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 102, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 104, or a variant thereofand/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:116, or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 118, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 120, or a variant thereofand/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:132, or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 134, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 136, or a variant thereofand/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:140, or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 142, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 144, or a variant thereofand/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:156, or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 158, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 160, or a variant thereofand/or CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO:172, or a variant thereof; CDR-H2 comprising the amino acid sequence setforth in SEQ ID NO: 174, or a variant thereof; and CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 176, or a variant thereofand/or a light chain immunoglobulin or variable region thereof thatcomprises: CDR-L1 comprising the amino acid sequence set forth in SEQ IDNO: 12, or a variant thereof; CDR-L2 comprising the amino acid sequenceset forth in SEQ ID NO: 14, or a variant thereof; and CDR-L3 comprisingthe amino acid sequence set forth in SEQ ID NO: 16, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:28, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 30, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 32, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:44, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 46, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 48, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:60, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 62, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 64, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:76, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 78, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 80, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:92, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 94, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 96, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:108, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 110, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 112, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:124, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 126, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 128, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:148, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 150, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 152, or a variant thereofand/or CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO:164, or a variant thereof; CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 166, or a variant thereof; and CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 168, or a variant thereof.6. The antibody or antigen-binding fragment thereof of claim 1comprising: (1) a heavy chain immunoglobulin or variable region thereofcomprising a CDR-H1 comprising the amino acid sequence set forth in SEQID NO: 4, or a variant thereof; a CDR-H2 comprising the amino acidsequence set forth in SEQ ID NO: 6, or a variant thereof; and a CDR-H3comprising the amino acid sequence set forth in SEQ ID NO: 8, or avariant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 12, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 14, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 16, ora variant thereof; (2) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 20, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 22, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 24, ora variant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 28, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 30, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 32, ora variant thereof; (3) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 36, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 38, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 40, ora variant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 44, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 46, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 48, ora variant thereof; (4) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 52, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 54, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 56, ora variant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 60, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 62, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 64, ora variant thereof; (5) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 68, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 70, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 72, ora variant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 76, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 78, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 80, ora variant thereof; (6) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 84, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 86, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 88, ora variant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 92, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 94, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 96, ora variant thereof; (7) a heavy chain immunoglobulin or variable regionthereof comprising a CDR-H1 comprising the amino acid sequence set forthin SEQ ID NO: 100, or a variant thereof; a CDR-H2 comprising the aminoacid sequence set forth in SEQ ID NO: 102, or a variant thereof; and aCDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 104,or a variant thereof; and a light chain immunoglobulin or variableregion thereof comprising a CDR-L1 comprising the amino acid sequenceset forth in SEQ ID NO: 108, or a variant thereof; a CDR-L2 comprisingthe amino acid sequence set forth in SEQ ID NO: 110, or a variantthereof; and a CDR-L3 comprising the amino acid sequence set forth inSEQ ID NO: 112, or a variant thereof; (8) a heavy chain immunoglobulinor variable region thereof comprising a CDR-H1 comprising the amino acidsequence set forth in SEQ ID NO: 116, or a variant thereof; a CDR-H2comprising the amino acid sequence set forth in SEQ ID NO: 118, or avariant thereof; and a CDR-H3 comprising the amino acid sequence setforth in SEQ ID NO: 120, or a variant thereof; and a light chainimmunoglobulin or variable region thereof comprising a CDR-L1 comprisingthe amino acid sequence set forth in SEQ ID NO: 124, or a variantthereof; a CDR-L2 comprising the amino acid sequence set forth in SEQ IDNO: 126, or a variant thereof; and a CDR-L3 comprising the amino acidsequence set forth in SEQ ID NO: 128, or a variant thereof; (9) a heavychain immunoglobulin or variable region thereof comprising a CDR-H1comprising the amino acid sequence set forth in SEQ ID NO: 132, or avariant thereof; a CDR-H2 comprising the amino acid sequence set forthin SEQ ID NO: 134, or a variant thereof; and a CDR-H3 comprising theamino acid sequence set forth in SEQ ID NO: 136, or a variant thereof;and a light chain immunoglobulin or variable region thereof comprising aCDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 124,or a variant thereof; a CDR-L2 comprising the amino acid sequence setforth in SEQ ID NO: 126, or a variant thereof; and a CDR-L3 comprisingthe amino acid sequence set forth in SEQ ID NO: 128, or a variantthereof; (10) a heavy chain immunoglobulin or variable region thereofcomprising a CDR-H1 comprising the amino acid sequence set forth in SEQID NO: 140, or a variant thereof; a CDR-H2 comprising the amino acidsequence set forth in SEQ ID NO: 142, or a variant thereof; and a CDR-H3comprising the amino acid sequence set forth in SEQ ID NO: 144, or avariant thereof; and a light chain immunoglobulin or variable regionthereof comprising a CDR-L1 comprising the amino acid sequence set forthin SEQ ID NO: 148, or a variant thereof; a CDR-L2 comprising the aminoacid sequence set forth in SEQ ID NO: 150, or a variant thereof; and aCDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 152,or a variant thereof; (11) a heavy chain immunoglobulin or variableregion thereof comprising a CDR-H1 comprising the amino acid sequenceset forth in SEQ ID NO: 156, or a variant thereof; a CDR-H2 comprisingthe amino acid sequence set forth in SEQ ID NO: 158, or a variantthereof; and a CDR-H3 comprising the amino acid sequence set forth inSEQ ID NO: 160, or a variant thereof; and a light chain immunoglobulinor variable region thereof comprising a CDR-L1 comprising the amino acidsequence set forth in SEQ ID NO: 164, or a variant thereof; a CDR-L2comprising the amino acid sequence set forth in SEQ ID NO: 166, or avariant thereof; and a CDR-L3 comprising the amino acid sequence setforth in SEQ ID NO: 168, or a variant thereof; or (12) a heavy chainimmunoglobulin or variable region thereof comprising a CDR-H1 comprisingthe amino acid sequence set forth in SEQ ID NO: 172, or a variantthereof; a CDR-H2 comprising the amino acid sequence set forth in SEQ IDNO: 174, or a variant thereof; and a CDR-H3 comprising the amino acidsequence set forth in SEQ ID NO: 176, or a variant thereof; and a lightchain immunoglobulin or variable region thereof comprising a CDR-L1comprising the amino acid sequence set forth in SEQ ID NO: 124, or avariant thereof; a CDR-L2 comprising the amino acid sequence set forthin SEQ ID NO: 126, or a variant thereof; and a CDR-L3 comprising theamino acid sequence set forth in SEQ ID NO: 128, or a variant thereof.7. The antigen-binding protein of claim 1 comprising: (a) a heavy chainimmunoglobulin or variable region thereof that comprises the amino acidsequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130,138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or224, or a variant thereof; and/or (b) a light chain immunoglobulin orvariable region thereof that comprises the amino acid sequence set forthin SEQ ID NO: 10, 26, 42, 58, 74, 90, 106, 122, 146, 162, 182, 186, 190,194, 198, 202, 206, 210, 214, 218, 222 or 226, or a variant thereof. 8.An isolated antibody or antigen-binding fragment thereof comprising: (a)a heavy chain immunoglobulin variable region that comprises the aminoacid sequence set forth in SEQ ID NO: 2, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 10; (b) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:18, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 26; (c) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 34, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:42; (d) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 50, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 58; (e) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:66, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 74; (f) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 82, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:90; (g) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 98, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 106; (h) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:114, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 122; (i) a heavy chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 130, and a light chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:122; (j) a heavy chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 138, and a light chainimmunoglobulin variable region that comprises the amino acid sequenceset forth in SEQ ID NO: 146; (k) a heavy chain immunoglobulin variableregion that comprises the amino acid sequence set forth in SEQ ID NO:154, and a light chain immunoglobulin variable region that comprises theamino acid sequence set forth in SEQ ID NO: 162; and/or (l) a heavychain immunoglobulin variable region that comprises the amino acidsequence set forth in SEQ ID NO: 170, and a light chain immunoglobulinvariable region that comprises the amino acid sequence set forth in SEQID NO:
 122. 9. The antibody or antigen-binding fragment thereof of claim8 wherein the heavy chain immunoglobulin variable region is linked to anIgG, IgG1 or IgG4 heavy chain constant region and the light chainimmunoglobulin variable region is linked to a kappa or lambda lightchain constant region.
 10. The antibody or antigen-binding fragmentthereof of claim 8 comprising: (a) a heavy chain immunoglobulin thatcomprises the amino acid sequence set forth in SEQ ID NO: 180, and alight chain immunoglobulin that comprises the amino acid sequence setforth in SEQ ID NO: 182; (b) a heavy chain immunoglobulin that comprisesthe amino acid sequence set forth in SEQ ID NO: 184, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 186; (c) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 188, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 190; (d) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 192, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 194; (e) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 196, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 198; (f) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 200, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 202; (g) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 204, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 206; (h) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 208, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 210; (i) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 212, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 214; (j) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 216, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 218; (k) a heavy chain immunoglobulin that comprises the aminoacid sequence set forth in SEQ ID NO: 220, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO: 222; and/or (l) a heavy chain immunoglobulin that comprises theamino acid sequence set forth in SEQ ID NO: 224, and a light chainimmunoglobulin that comprises the amino acid sequence set forth in SEQID NO:
 226. 11. The antibody or antigen-binding fragment thereof ofclaim 8 which is an antibody.
 12. The antibody or antigen-bindingfragment thereof of claim 8 which is multispecific.
 13. The antibody orantigen-binding fragment thereof of claim 1 which comprises one or moreof the following properties: Binds to human IL36R (IL-1RL2) with a K_(D)of about 2.18 nM to about 13.9 nM at 25° C. or with a K_(D) of about4.25 nM to about 29.5 nM at 37° C.; Binds to Macaca fascicularis IL36R(IL-1RL2) with a K_(D) of about 7.87 nM to about 34.4 nM at 25° C. orwith a K_(D) of about 14.4 nM to about 58.2 nM at 37° C.; Binds to humanIL36R (IL-1RL2) fused to a mouse IgG2a with a K_(D) of about 173 pM toabout 5.79 nM at 25° C. or with a K_(D) of about 205 pM to about 28.7 nMat 37° C.; Binds to human IL36R (IL-1RL2) fused to IL1RAcP extracellulardomain expressed with mouse IgG2a Fc tag with a K_(D) of about 212 pM toabout 14 nM at 25° C. or with a K_(D) of about 264 pM to about 40.9 nMat 37° C.; Competes with H4H14699P2; H4H14700P2; H4H14706P2; H4H14708P2;H4H14709P; H4H14728P; H4H14731P; H4H14732P2; H4H14734P2; H4H14757P;H4H14758P or H4H14760P2 for binding to IL36R (IL-1RL2); optionally withthe proviso that said antigen-binding protein is not APE6155 or anantigen-binding fragment thereof; Blocks activation of one or more NFκBelements, which is/are fused to a reporter gene, in a host cell, byIL-36R (IL-1RL2) in the presence of IL-1RAcP and IL36R ligand; Preventsor ameliorates skin inflammation or reduces skin thickness or totalpathology score or reduces pro-inflammatory cytokine levels in a subjectsuffering from skin inflammation; Prevents or ameliorates colitis orcolon inflammation or reduces fecal levels of LCN2 polypeptide in asubject with such colitis or inflammation Protects residues (a) 113-119,113-122, 116-119 and/or 116-122; and/or (b) 264-271, 267-271, 268-271,268-276, 268-277 and/or 271-276, of human IL36R (IL-1RL2) comprising theamino acid sequence set forth in SEQ ID NO: 227, when bound, fromdigestion with pepsin and/or Protease XIII and/or deuteration in thepresence of deuterium; Binds to IL36R (IL-1RL2) comprising the aminoacid sequence set forth in SEQ ID NO: 227 at residues 113-119, 113-122,116-119, 116-122, 264-271, 267-271, 268-271, 268-276, 268-277 and/or271-276; Binds Domain II of IL36R (IL-1RL2); Binds a polypeptidecomprising the amino acid sequence YKQILHLGKD (SEQ ID NO: 229) (aminoacids 113-122 of SEQ ID NO: 227); Inhibits IL36α, IL36β and/or IL36γ inin vitro epidermal keratinocytes, intestinal myofibroblasts and/or CD14+monocytes, with an IC₅₀ of about 1-6 nM; and/or Competitively inhibitsIL36α, IL36β and/or IL36γ-mediated activation of NFκB by IL36R.
 14. Acomplex comprising an antibody or antigen-binding fragment thereof ofclaim 1 bound to a IL36R polypeptide.
 15. A method for making anantibody or antigen-binding fragment thereof of claim 1 or animmunoglobulin chain thereof comprising: (a) introducing one or morepolynucleotides encoding an immunoglobulin chain of said antibody orantigen-binding fragment thereof into a host cell; (b) culturing thehost cell in a medium under conditions favorable to expression of thepolynucleotide; and (c) optionally, isolating the antibody orantigen-binding fragment thereof or immunoglobulin chain from the hostcell and/or medium in which the host cell is grown.
 16. The method ofclaim 15 wherein the host cell is a Chinese hamster ovary cell.
 17. Anantibody or antigen-binding fragment thereof or immunoglobulin chainwhich is a product of the method of claim
 15. 18. A polypeptidecomprising: (a) CDR-H1, CDR-H2, and CDR-H3 of a heavy chainimmunoglobulin or variable region thereof that comprises the amino acidsequence set forth in SEQ ID NO: 2, 18, 34, 50, 66, 82, 98, 114, 130,138, 154, 170, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220 or224, or a variant thereof; and/or (b) CDR-L1, CDR-L2, and CDR-L3 of alight chain immunoglobulin or variable region thereof that comprises theamino acid sequence set forth in SEQ ID NO: 10, 26, 42, 58, 74, 90, 106,122, 146, 162, 182, 186, 190, 194, 198, 202, 206, 210, 214, 218, 222 or226, or a variant thereof; or, (c) the amino acid sequence set forth ina member selected from the group consisting of SEQ ID NO: 1-226, or avariant thereof.
 19. A polynucleotide encoding the polypeptide of claim18.
 20. A vector comprising the polynucleotide of claim
 19. 21. A hostcell comprising the vector of claim
 20. 22. A composition or kitcomprising an antibody or antigen-binding fragment thereof of claim 1,optionally in association with a further therapeutic agent.
 23. Apharmaceutical composition comprising the antibody or antigen-bindingfragment thereof of claim 1 and a pharmaceutically acceptable carrierand, optionally, a further therapeutic agent.
 24. The pharmaceuticalcomposition of claim 23 in association with a further therapeutic agentwhich is an anti-inflammatory agent.
 25. The pharmaceutical compositionof claim 23 in association with a further therapeutic agent, wherein thefurther therapeutic agent is a member selected from the group consistingof an anti-TNFalpha antibody or antigen-binding fragment thereof, one ormore human TNF receptors or fragments thereof linked to animmunoglobulin, an IL17 inhibitor, an IL23p19 inhibitor, an IL12p40inhibitor, guselkumab, ustekinumab, brodalumab, ixekizumab, secukinumab,infliximab, adalimumab, etanercept, dupilumab, sarilumab, tocilizumab,golimumab, abatacept, tofacitinib, abatacept, a non-steroidalanti-inflammatory drug (NSAID), ibuprofen, naproxen, acetaminophen,aspirin, celecoxib, cyclophosphamide, methotrexate, a corticosteroid,cortisone and prednisone.
 26. A vessel or injection device comprisingthe antibody or antigen-binding fragment thereof of claim 1 or apharmaceutical composition comprising said antibody or fragment and apharmaceutically acceptable carrier.
 27. A method for treating orpreventing an IL-36R-mediated disease in a subject in need thereof,comprising administering, to the subject, a therapeutically effectiveamount of antibody or antigen-binding fragment thereof of claim 1optionally in association with a further therapeutic agent.
 28. A methodfor treating or preventing: an inflammatory disorder, an autoimmunedisorder, deficiency of interleukin IL-36 receptor antagonist (DITRA)syndrome, impetigo herpetiformis, acrodermatitis, a skin neutrophilicpustular disease, a pustular disease, psoriasis, generalized pustularpsoriasis, psoriasis vulgaris, palmoplantar pustular psoriasis,palmoplantar pustulosis, colitis, airway inflammation, jointinflammation, kidney inflammation, alopecia areata, skin inflammation,acanthosis, hyperkeratosis, kindler syndrome, systemic lupuserythematosus (SLE), nephrotic syndrome, ANCA (anti-neutrophilcytoplasmic antibody)-associated vasculopathies, tubulointerstitiallesions or glomerulonephritis; comprising administering, to the subject,a therapeutically effective amount of antibody or antigen-bindingfragment thereof of claim 1 optionally in association with a furthertherapeutic agent.
 29. The method of claim 27, for treating orpreventing psoriasis or inflammatory bowel disease.
 30. A method foradministering an antibody or antigen-binding fragment thereof of claim 1to a subject comprising injecting the antibody or antigen-bindingfragment thereof into the body of the subject, optionally in associationwith a further therapeutic agent.
 31. The method of claim 30 wherein theantibody or antigen-binding fragment thereof is injected into the bodyof the subject subcutaneously, intravenously or intramuscularly.
 32. Themethod of claim 27 wherein the subject has a homozygous or heterozygousIL36RN mutation genotype.